Rock kinase inhibitors

ABSTRACT

The present invention relates to compounds that inhibit ROCK activity. In particular, the present invention relates to compounds, pharmaceutical compositions and methods of use, such as methods of inhibiting ROCK activity and methods for treating, for example cerebral cavernous malformation syndrome (CCM) and cardiovascular diseases using the compounds and pharmaceutical compositions of the present invention.

FIELD OF THE INVENTION

The present invention relates to compounds that are Rho-associatedprotein kinases (ROCK) inhibitors. In particular, the present inventionrelates to compounds that selectively inhibit the activity of ROCK,pharmaceutical compositions comprising the compounds and methods of usetherefor.

BACKGROUND OF THE INVENTION

ROCK is is a critical RhoA effector involved in regulation of multiplecellular signaling pathways through phosphorylation of multiplesubstrates, including myosin light chain (MLC), LIM Kinase, MYPT1, andCPI-17. ROCK substrates are involved in smooth muscle contractility,vascular permeability, actin filament organization, cell adhesion, cellmigration, growth control and cytokinesis. The direct involvement ofROCK in smooth muscle contractility results from increasedphosphorylation of both MLC, which increases contractility uponphosphorylation, and MLC-phosphatase, which is inhibited uponphosphorylation, thereby further increaser MLC phosphorylation ROCKactivation, thus, results in increased smooth muscle contractility.

Literature reports suggest a link between ROCK activity and numerouscardiovascular diseases associated with vasotonic dysfunction, includinghypertension, atherosclerosis, ischemic stroke, coronary vasospasm,cerebral vasospasm, angina, erectile dysfunction, and renal disease andalso a link with other smooth muscle hyper-reactivity, such as glaucomaand asthma.

Cerebral cavernous malformation (CCM) syndrome is a disease associatedwith defective endothelial junctions predominantly in the brainpredisposing patients to a lifetime risk of seizures, hemorrhagicstroke, and other neurological deficits. Cerebral cavernousmalformations (CCMs) are vascular malformations mostly found within thecentral nervous system, e.g., brain stem and spinal cord, that can occurin sporadic or autosomal dominant inherited forms, the latter of whichmap to three loci, KRIT-1/CCM1, MGC4607/OSM/CCM2, and PDCD10/CCM3 (e.g.,see Glading et al., (2007) J. Cell Biol. 179:247-254).

CCM1, CCM2 and CCM3 proteins each interact with a number of proteins,including Rap1, a RhoA-GTPase. Rap1 plays a role in regulating the actincytoskeleton by stimulating the formation of actin fibers. Inactivatingmutations in CCM1, CCM2 or CCM3 are sufficient to induce CCM vascularlesions by inhibiting endothelial cell vessel-like tube formation andextracellular matrix invasion leading to weakened junctions betweencells and increased leakage from blood vessels. The loss of expressionof the CCM1, -2, or -3 proteins causes a marked overexpression andincreased activation of Rap1. Increased Rap1 activation is associatedwith increased Rho kinase-dependent (ROCK) phosphorylation, and shorthairpin RNA knockdown of ROCK rescued endothelial cells from CCMpathology (e.g., see Borikova et al., (2010) J. Biol. Chem.285:11760-11764).

CCM patients unfortunately have limited treatment options and there areno approved drugs in the United States for treating CCM. Fasudilhydrochloride, an isoquinoline sulfonamide derivative, is a potent ROCKinhibitor that has been approved outside the United States for treatingcerebral vasospasm after subarachnoid hemorrhage and symptoms ofcerebral ischemia. Fasudil has been used off-label by physicians totreat CCM, presumably by inhibiting ROCK kinase activity; however,Fasudil, which shows poor selectivity over other AGC-family kinases,also exhibits a number of serious toxic side effects and drawbacksincluding mandated short-course treatments, low oral bioavailability,cell toxicity and blood pressure fluctuations (e.g., see Xin et al.,(2015) BioSci Rep. 35: 1-13).

There is a serious unmet medical need to treat patients having CCM, andmore broadly for numerous other diseases involving elevated smoothmuscle contractility, such as cardiovascular disease, with prevailingevidence pointing towards selective ROCK kinase inhibitors as a likelysource for pharmacological therapeutic intervention. In conjunction,there is a need to develop new ROCK kinase inhibitors that have improvedtreatment times, improved oral bioavailability, reduced cell toxicityand reduced off-target activity, particularly over the AGC-family kinasemembers.

SUMMARY OF THE INVENTION

The present inventors recognized a need to develop new ROCK kinaseinhibitors that demonstrate improved cellular potency, kinomeselectivity, efficacy, stability and safety. The compounds andcompositions of the present invention advantageously overcome one ormore of these shortcomings by providing potent, selective and orallyactive ROCK inhibitors.

In one aspect of the invention, compounds are provided that inhibit ROCKkinase activity. In certain embodiments, the compounds are representedby formula (Ia), formula (Ib), formula (IIa) or formula (IIb):

or a pharmaceutically acceptable salt thereof:

wherein:

X is a partially saturated aza-containing heteroaryl;

Y is N or CH;

each R¹ is cyano, hydroxyl, hydroxyalkyl, halogen, haloalkyl, alkoxy,Q-C₁-C₃ alkyl, —N(R⁴)₂, —NR⁴C(O)C1-C3 alkyl or cycloalkyl;

Q is a bond, O or N;

each R² is hydrogen or halogen;

each R³ is hydrogen, halogen or C₁-C₃ alkyl;

each R⁴ is hydrogen or C₁-C₃ alkyl; and

m is 0, 1, 2 or 3.

In another aspect of the invention, pharmaceutical compositions areprovided comprising a therapeutically effective amount of a compound offormula (Ia), formula (Ib), formula (IIa) or formula (IIb) andpharmaceutically acceptable excipients, salts, solvates, carriers and/ordiluents.

In yet another aspect, the present invention is directed to a method oftreating a cardiovascular disease associated with increased vasotensioncomprising administering a therapeutically effective amount of acompound of formula (Ia), formula (Ib), formula (IIa) or formula (IIb)or pharmaceutical composition thereof to a patient in need thereof. Inone embodiment, the cardiovascular disease is hypertension,atherosclerosis, ischemic stroke, coronary vasospasm, cerebralvasospasm, angina and erectile dysfunction.

In yet another aspect, the present invention is directed to a method oftreating diseases involving elevated non-vascular smooth musclecontractility comprising administering a therapeutically effectiveamount of a compound of formula (Ia), formula (Ib), formula (IIa) orformula (IIb) or pharmaceutical composition thereof to a patient in needthereof. In one embodiment, the disease involving elevated non-vascularsmooth muscle contractility asthma and glaucoma.

In yet another aspect of the invention, methods for inhibiting ROCKactivity in a cell comprising contacting the cell with a compound offormula (Ia), formula (Ib), formula (IIa) or formula (IIb) are provided.Also provided are methods for treating CCM in a patient comprisingadministering a therapeutically effective amount of a compound offormula (Ia), formula (Ib), formula (IIa) or formula (IIb) orpharmaceutical composition thereof to a patient in need thereof.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to ROCK kinase inhibitors. In particular,the present invention relates to compounds that selectively inhibit theactivity of a ROCK kinase, pharmaceutical compositions comprising atherapeutically effective amount of the compounds, and methods of usetherefor, such as methods for treating cerebral cavernous malformationsyndrome (CCM) and cardiovascular diseases.

Definitions

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as is commonly understood by one skilled in theart to which this invention belongs. All patents, patent applications,and publications referred to herein are incorporated by reference to theextent they are consistent with the present disclosure. Terms and rangeshave their generally defined definition unless expressly definedotherwise.

For simplicity, chemical moieties are defined and referred to throughoutprimarily as univalent chemical moieties (e.g., alkyl, aryl, etc.).Nevertheless, such terms may also be used to convey correspondingmultivalent moieties under the appropriate structural circumstancesclear to those skilled in the art. All atoms are understood to havetheir normal number of valences for bond formation (i.e., 4 for carbon,3 for N, 2 for O, and 2, 4, or 6 for S, depending on the oxidation stateof the S).

As used herein, a “ROCK kinase” refers to a member of the Rho-associatedprotein kinase (“ROCK”) family including ROCK1 and ROCK2 kinases.

As used herein, an “ROCK inhibitor” refers to compounds of the presentinvention that are represented by formulae (I) or (II) as describedherein. These compounds are capable of negatively modulating orinhibiting all or a portion of the enzymatic activity of a ROCK kinase.

The term “alkyl” as employed herein refers to straight and branchedchain aliphatic groups having from 1 to 12 carbon atoms. As such,“alkyl” encompasses C₁, C₂, C₃, C₄, C₅, C₆, C₇, C₈, C₉, C₁₀, C₁₁ and C₁₂groups. Examples of alkyl groups include, without limitation, methyl,ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl,pentyl, and hexyl.

The term “cyano” refers to —CN.

The term “cycloalkyl” as employed herein is a saturated and partiallyunsaturated cyclic hydrocarbon group having 3 to 12 carbons. As such,“cycloalkyl” includes C₃, C₄, C₅, C₆, C₇, C₈, C₉, C₁₀, C₁₁ and C₁₂cyclic hydrocarbon groups. Examples of cycloalkyl groups include,without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl,cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl.

As used herein, the term “heteroaryl” refers to a group having 5 to 14ring atoms, preferably 5, 6, 10, 13 or 14 ring atoms; having 6, 10, or14 it electrons shared in a cyclic array; and having, in addition tocarbon atoms, from one to three heteroatoms that are each independentlyN, O, or S. “Heteroaryl” also includes fused multicyclic (e.g.,bicyclic) ring systems in which one or more of the fused rings isnon-aromatic, provided that at least one ring is aromatic and at leastone ring contains an N, O, or S ring atom.

Examples of heteroaryl groups include acridinyl, azocinyl,benzimidazolyl, benzofuranyl, benzo[d]oxazol-2(3H)-one,2H-benzo[b][1,4]oxazin-3(4H)-one, benzothiofuranyl, benzothiophenyl,benzoxazolyl, benzthiazolyl, benztriazolyl, benztetrazolyl,benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl,4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl, furanyl,furazanyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl,indolizinyl, indolyl, 3H-indolyl, isobenzofuranyl, isochromanyl,isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl,isoxazolyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl,1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl,1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl, pyrimidinyl,phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl,phenoxathiinyl, phenoxazinyl, phthalazinyl, piperonyl, pteridinyl,purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl,pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl,pyridyl, pyrimidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl,quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl,tetrahydroisoquinolinyl, tetrahydroquinolinyl, tetrazolyl,6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl,1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl,thienyl, thienothiazolyl, thienooxazolyl, thienoimidazolyl, thiophenyl,triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl,1,3,4-triazolyl, and xanthenyl.

The term “aza-containing heteroaryl” refers to a heteroaryl group thatcontains a nitrogen atom in place of a carbon atom in a monocyclic,bicyclic, polycyclic or fused group having 5 to 14 ring atoms,preferably 5, 6, 10, 13 or 14 ring atoms; having 6, 10, or 14 itelectrons shared in a cyclic array. “Aza-containing heteroaryl” alsoincludes fused multicyclic (e.g., bicyclic) ring systems in which one ormore of the fused rings is non-aromatic, provided that at least one ringis aromatic and at least one ring contains an N.

Exemplary aza-containing heteroaryl groups include, but are not limitedto, indolizinyl, indolyl, isoindolyl, 3H-indolyl, indolinyl, indazolyl,1H-indazolyl, benzimidazolyl, benzthiazolyl, purinyl, 4H-quinolizinyl,quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl,quinoxalinyl, 1,8-naphthyridinyl, pteridinyl, quinuclidinyl, carbazolyl,acridinyl, phenazimyl, phenothiazinyl, phenoxazinyl, 2H-pyrrolyl,pyrrolyl, 2-pyrrolinyl, 3-pyrrolinyl, oxazolyl, thiazolyl, imidazolyl,2-imidazolinyl, pyrazolyl, 2-pyrazolinyl, isoxazolyl, isothiazolyl,1,2,3-oxadiazolyl, 1,2,3-triazolyl, 1,3,4-thiadiazolyl, pyridyl,pyridazinyl, pyrimidinyl, and 1,3,5-triazinyl.

The term “haloalkyl” refers to an alkyl chain in which one or morehydrogens have been replaced by a halogen. Exemplary haloalkyls aretrifluoromethyl, difluoromethyl, flurochloromethyl, and fluoromethyl.

The term “halogen” or “halo” as employed herein refers to chlorine,bromine, fluorine, or iodine.

The term “hydroxyl” refers to —OH.

The term “hydroxyalkyl” refers to -alkyl-OH.

As used herein, “an effective amount” of a compound is an amount that issufficient to negatively modulate or inhibit the activity of a ROCKkinase.

As used herein, a “therapeutically effective amount” of a compound is anamount that is sufficient to ameliorate or in some manner reduce asymptom or stop or reverse progression of a condition, e.g., CCM, ornegatively modulate or inhibit the activity of a ROCK kinase. Suchamount may be administered as a single dosage or may be administeredaccording to a regimen, whereby it is effective.

As used herein, “treatment” means any manner in which the symptoms orpathology of a condition, disorder or disease in a patient areameliorated or otherwise beneficially altered.

As used herein, amelioration of the symptoms of a particular disorder byadministration of a particular compound or pharmaceutical compositionrefers to any lessening, whether permanent or temporary, lasting ortransient, that can be attributed to or associated with administrationof the composition.

Compounds

In one aspect of the invention, compounds are provided represented byformula (Ia), formula (Ib), formula (IIa) or formula (IIb):

or a pharmaceutically acceptable salt thereof:

wherein:

X is a partially saturated aza-containing heteroaryl;

Y is N or CH;

each R¹ is cyano, hydroxyl, hydroxyalkyl, halogen, haloalkyl, alkoxy,Q-C₁-C₃ alkyl, —N(R⁴)₂, —NR⁴C(O)C1-C3 alkyl or cycloalkyl;

Q is a bond, O or N;

each R² is hydrogen or halogen;

each R³ is hydrogen, halogen or C₁-C₃ alkyl;

each R⁴ is hydrogen or C₁-C₃ alkyl; and

m is 0, 1, 2 or 3.

In one aspect of the invention, X is an aza-containing heteroarylselected from indolizinyl, indolyl, isoindolyl, 3H-indolyl, indolinyl,indazolyl, 1H-indazolyl, benzimidazolyl, benzthiazolyl, purinyl,4H-quinolizinyl, quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl,quinazolinyl, quinoxalinyl, 1,8-naphthyridinyl, pteridinyl,quinuclidinyl, carbazolyl, acridinyl, phenazimyl, phenothiazinyl,phenoxazinyl, 2H-pyrrolyl, pyrrolyl, 2-pyrrolinyl, 3-pyrrolinyl,oxazolyl, thiazolyl, imidazolyl, 2-imidazolinyl, pyrazolyl,2-pyrazolinyl, isoxazolyl, isothiazolyl, 1,2,3-oxadiazolyl,1,2,3-triazolyl, 1,3,4-thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl,and 1,3,5-triazinyl. In one embodiment, X is an R¹-substitutedindolin-1-yl.

In one embodiment, X is:

wherein m, Y and R¹ are as defined herein.

In one embodiment, m is one and R¹ is cyano, hydroxyl, hydroxyalkyl,halogen, haloalkyl, alkoxy, Q-C₁-C₃ alkyl, —N(R⁴)₂, —NR⁴C(O)C1-C3 alkylor cycloalkyl.

In one embodiment, R¹ is cyano. In one embodiment, R¹ is hydroxyl. Inone embodiment, R¹ is halogen. In one embodiment, the halogen ischlorine or fluorine. In one embodiment, R¹ is haloalkyl. In oneembodiment, the haloalkyl is trifluoromethyl. In one embodiment, R¹ isQ-C₁-C₃ alkyl. In one embodiment, Q is a bond and the C₁-C₃ alkyl ismethyl, ethyl or isopropyl. In one embodiment, R¹ is Q-C₁-C₃ alkyl,wherein Q is O and the C₁-C₃ alkyl is methyl, ethyl or isopropyl. In oneembodiment, R¹ is Q-C₁-C₃ alkyl, wherein Q is N and the C₁-C₃ alkyl ismethyl, ethyl or isopropyl. In one embodiment, R¹ is hydroxyalkyl,wherein the hydroxyalkyl is hydroxymethyl. In one embodiment, R¹ isalkoxy. In one embodiment, the alkoxy is methoxy. In one embodiment, R¹is the —N(R⁴)₂. In one embodiment, each R⁴ group is hydrogen. In oneembodiment, each R⁴ group is C₁-C₃ alkyl. In one embodiment, R¹ is—NR⁴C(O)C1-C3 alkyl. In one embodiment, the R⁴ group is hydrogen and theC₁-C₃ alkyl is methyl. In one embodiment, R¹ is cycloalkyl, wherein thecycloalkyl is cyclopropyl.

In one embodiment, m is two and each R¹ is independently cyano,hydroxyl, hydroxyalkyl, halogen, haloalkyl, alkoxy, Q-C₁-C₃ alkyl,—N(R⁴)₂, —NR⁴C(O)C1-C3 alkyl or cycloalkyl. In certain embodiments, eachR¹ is halogen. In one embodiment, one R¹ is cyano and the other R¹ ishydroxyl, hydroxyalkyl, halogen, haloalkyl, alkoxy, Q-C₁-C₃ alkyl,—N(R⁴)₂, —NR⁴C(O)C1-C3 alkyl or cycloalkyl.

In one embodiment, X is:

wherein each R¹ is hydrogen or methyl. In one embodiment, each R¹ ismethyl.

In one embodiment, R² is halogen. In certain embodiments, the halogen ischlorine.

In one embodiment, R³ is halogen or C₁-C₃ alkyl. In certain embodiments,the halogen is fluorine. In other embodiments, R³ is C₁-C₃ alkyl,wherein the C₁-C₃ alkyl is methyl.

In certain embodiments, compounds of formula (Ia), formula (Ib), formula(IIa) or formula (IIb) are:

and pharmaceutically acceptable salts of the foregoing compounds.

The compounds of formula (Ia), formula (Ib), formula (IIa) or formula(IIb) may be formulated into pharmaceutical compositions.

The compounds disclosed herein include all pharmaceutically acceptableisotopically-labelled compounds, in which one or more atoms of thecompounds disclosed herein are replaced by atoms having the same atomicnumber and an atomic mass or mass number different from the atomic massor mass number usually found in nature. Examples of isotopes that can beincorporated into the compounds disclosed herein include isotopes ofhydrogen, carbon, nitrogen, oxygen, phosphorous, sulfur, fluorine,chlorine, and iodine, such as ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ¹³N, ¹⁵N, ¹⁵O, ¹⁷O,³¹P, ³²P, ³⁵S, ¹⁸F, ³⁶Cl, ¹²³I, and ¹²⁵I. Such radiolabeled compoundscould be useful to help determine or measure the effectiveness of thecompounds, for example, by characterizing binding affinities andkinetics of binding to ROCK and other modes of action. Certain compoundsof the disclosure, when labelled with radioactive isotopes, such astritium, ³H, and carbon-14, ¹⁴C, could also be useful in characterizingdrug distribution and in vivo tissue expression of ROCK.

When labelled with heavier isotopes such as deuterium, ²H, compoundsdisclosed herein may exhibit therapeutic advantages such as greatermetabolic stability, resulting in longer in-vivo half-lives, which inturn could reduce the therapeutic dosing requirements.

In order to better characterize substrate receptor occupancy or tissueexpression in live specimens statically or longitudinally, replacingatoms of compounds disclosed herein with positron emitting isotopes,such as ¹¹C, ¹⁴F, ¹⁵O and ¹³N, may allow for enabling Positron EmissionTomography (PET) studies.

Isotopically-labeled compounds as disclosed herein can be prepared byconventional techniques known to those skilled in the art or byprocesses that are analogous to those described in the associatedexamples and schemes using an appropriate isotopically-labeled reagentin place of the non-labeled reagent previously employed.

Some of the compounds disclosed herein may exist as stereoisomers. Thecompounds disclosed herein include all stereoisomers, both as pureindividual stereoisomer preparations as well as preparations enriched ofeach, and both the racemic mixures of such stereoisomers and theindividual diasteromers and enantiomers that may be separated accordingto methods known to those skilled in the art. Additionally, thecompounds disclosed herein include all individual tautomeric sates ofthe compounds and mixtures thereof.

Pharmaceutical Compositions

In another aspect, the invention provides pharmaceutical compositionscomprising a ROCK kinase inhibitor according to the invention and apharmaceutically acceptable carrier, excipient, or diluent. Compounds ofthe invention may be formulated by any method well known in the art andmay be prepared for administration by any route, including, withoutlimitation, parenteral, oral, sublingual, transdermal, topical,subcutaneous, intranasal, intratracheal, or intrarectal. In certainembodiments, compounds of the invention are administered intravenouslyin a hospital setting. In certain other embodiments, administration maypreferably be by the oral route.

The characteristics of the carrier will depend on the route ofadministration. As used herein, the term “pharmaceutically acceptable”means a non-toxic material that is compatible with a biological systemsuch as a cell, cell culture, tissue, or organism, and that does notinterfere with the effectiveness of the biological activity of theactive ingredient(s). Thus, compositions according to the invention maycontain, in addition to the inhibitor, diluents, fillers, salts,buffers, stabilizers, solubilizers, and other materials well known inthe art. The preparation of pharmaceutically acceptable formulations isdescribed in, e.g., Remington's Pharmaceutical Sciences, 18th Edition,ed. A. Gennaro, Mack Publishing Co., Easton, Pa., 1990.

As used herein, the term pharmaceutically acceptable salts refers tosalts that retain the desired biological activity of theabove-identified compounds and exhibit minimal or no undesiredtoxicological effects. Examples of such salts include, but are notlimited to acid addition salts formed with inorganic acids (for example,hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid,nitric acid, and the like), and salts formed with organic acids such asacetic acid, oxalic acid, tartaric acid, succinic acid, malic acid,ascorbic acid, benzoic acid, tannic acid, pamoic acid, alginic acid,polyglutamic acid, naphthalenesulfonic acid, naphthalenedisulfonic acid,and polygalacturonic acid. The compounds can also be administered aspharmaceutically acceptable quaternary salts known by those skilled inthe art, which specifically include the quaternary ammonium salt of theformula —NR+Z—, wherein R is hydrogen, alkyl, or benzyl, and Z is acounterion, including chloride, bromide, iodide, —O-alkyl,toluenesulfonate, methylsulfonate, sulfonate, phosphate, or carboxylate(such as benzoate, succinate, acetate, glycolate, maleate, malate,citrate, tartrate, ascorbate, benzoate, cinnamoate, mandeloate,benzyloate, and diphenylacetate).

The active compound is included in the pharmaceutically acceptablecarrier or diluent in an amount sufficient to deliver to a patient atherapeutically effective amount without causing serious toxic effectsin the patient treated. A dose of the active compound for all of theabove-mentioned conditions is in the range from about 0.01 to 300 mg/kg,preferably 0.1 to 100 mg/kg per day, more generally 0.5 to about 25 mgper kilogram body weight of the recipient per day. A typical topicaldosage will range from 0.01-3% wt/wt in a suitable carrier. Theeffective dosage range of the pharmaceutically acceptable derivativescan be calculated based on the weight of the parent compound to bedelivered. If the derivative exhibits activity in itself, the effectivedosage can be estimated as above using the weight of the derivative, orby other means known to those skilled in the art.

The pharmaceutical compositions comprising compounds of the presentinvention may be used in the methods described herein.

Methods of Use

In yet another aspect, the invention provides for methods for inhibitingROCK activity in a cell, comprising contacting the cell in whichinhibition of ROCK activity is desired with a therapeutically effectiveamount of a compound of formula (Ia), formula (Ib), formula (IIa) orformula (IIb), pharmaceutically acceptable salts thereof orpharmaceutical compositions containing the compound or pharmaceuticallyacceptable salt thereof.

The compositions and methods provided herein are deemed particularlyuseful for inhibiting ROCK activity in a cell. In one embodiment, a cellin which inhibition of ROCK activity is desired is contacted with aneffective amount of a compound of formula (Ia), formula (Ib), formula(IIa) or formula (IIb) to negatively modulate the activity of a ROCKkinase. In other embodiments, a therapeutically effective amount ofpharmaceutically acceptable salt or pharmaceutical compositionscontaining the compound of formula (Ia), formula (Ib), formula (IIa) orformula (IIb) may be used. In certain embodiments, the contacting thecell with an effective amount or a therapeutically effective amount of acompound of formula (Ia), formula (Ib), formula (IIa) or formula (IIb)occurs in vivo. In certain embodiments, the contacting the cell with aneffective amount or a therapeutically effective amount of a compound offormula (Ia), formula (Ib), formula (IIa) or formula (IIb) occurs invitro. In one embodiment, the therapeutically effective amount of thecompound of formula (Ia), formula (Ib), formula (IIa) or formula (IIb)is between about 0.01 to 300 mg/kg per day. In one embodiment, thetherapeutically effective amount of the compound of formula (Ia),formula (Ib), formula (IIa) or formula (IIb) is between about 0.1 to 100mg/kg per day.

In one embodiment, the compound of formula (Ia), formula (Ib), formula(IIa) or formula (IIb) preferentially inhibits the activity of ROCK2kinase over ROCK1 kinase. In another embodiment, the compound of formula(Ia), formula (Ib), formula (IIa) or formula (IIb) inhibits the activityof ROCK2 kinase and ROCK1 kinase to a similar degree. In one embodiment,the therapeutically effective amount of the compound of formula (Ia),formula (Ib), formula (IIa) or formula (IIb) is between about 0.01 to300 mg/kg per day. In one embodiment, the therapeutically effectiveamount of the compound of formula (Ia), formula (Ib), formula (IIa) orformula (IIb) is between about 0.1 to 100 mg/kg per day.

In one embodiment, methods of for treating a patient having cerebralcavernous malformation syndrome (CCM), comprising administering to thepatient a therapeutically effective amount of a compound of formula(Ia), formula (Ib), formula (IIa) or formula (IIb), or apharmaceutically acceptable salt thereof, alone or combined with apharmaceutically acceptable carrier, exicipient or diluents areprovided.

In another embodiment, methods of treating a cardiovascular diseaseassociated with increased vasotension comprising administering atherapeutically effective amount of a compound of formula (Ia), formula(Ib), formula (IIa) or formula (IIb) or pharmaceutical compositionthereof to a patient in need thereof are provided. In one embodiment,the cardiovascular disease is hypertension, atherosclerosis, ischemicstroke, coronary vasospasm, cerebral vasospasm, angina and erectiledysfunction.

In embodiment, methods of treating diseases involving elevatednon-vascular smooth muscle contractility comprising administering atherapeutically effective amount of a compound of formula (Ia), formula(Ib), formula (IIa) or formula (IIb) or pharmaceutical compositionthereof to a patient in need thereof are provided. In one embodiment,the disease involving elevated non-vascular smooth muscle contractilityasthma and glaucoma.

By negatively modulating the activity of ROCK, particularly in cases forcells overexpressing the ROCK enzyme or autosomal dominant mutationsthat activate the ROCK enzyme, the methods are designed to modulate theactivity of ROCK, in some embodiments, to inhibit the formation of newCCMs and/or reduce or eradicate existing CCMs in treated patients ortreat certain cardiovascular diseases or diseases involving elevatednon-vascular smooth muscle contractility. The cells/patient may becontacted in a single dose or multiple doses in accordance with aparticular treatment regimen to effect the desired negative modulationof ROCK. For example, the size and number of CCMs may be monitored usingwell known methods, including CT scans or MRI scans, to assess theeffectiveness of treatment and dosages may be adjusted accordingly bythe attending medical practitioner.

The concentration and route of administration to the patient may varydepending on the severity of the disease. The compounds,pharmaceutically acceptable salts thereof and pharmaceuticalcompositions comprising such compounds and salts also may beco-administered with other compounds or used in combination with othertreatments, such as surgical intervention, either as an adjuvant priorto surgery or post-operatively. The degree of ROCK kinase inhibition maybe monitored in the patient using well known assay methods, includingthose in Example A, to access the effectiveness of treatment, along withother cranial imaging techniques and instrumentation, such as CT-scans,MRIs, X-rays, etc. and dosages may be adjusted accordingly by theattending medical practitioner.

Reaction Schemes and Examples

The compounds of the present invention may be prepared usingcommercially available reagents using the synthetic methods and reactionschemes described herein, or using other reagents and conventionalmethods well known to those skilled in the art.

For instance, compounds of the present invention may be preparedaccording to the general reaction schemes I, II, and III.

Scheme I illustrates the preparation of compounds of formula (I) of thepresent invention. An R²-substituted haloisoquinoline A undergoesaromatic electrophilic substitution reaction with sulfurochloridic acidunder elevated temperature to afford halosulfonated intermediate B (Step1). Reaction of halosulfonated intermediate B under acidic conditions,e.g., HCl, and elevated temperature affords sulfonic acid substitutedisoquinolinone C (Step 2). The sulfonic acid is converted to thesulfonyl chloride typically using sulfuryl chloride in DMF at reducedtemperatures (Step 3). The addition of R¹-substituted aza-containingheteroaryl X typically proceeds under basic conditions using anappropriate base such as TEA in a solvent such as DCM to affordcompounds E of formula (I) (Step 4).

Scheme II illustrates the preparation of compounds of formula (II) ofthe present invention, wherein R³ is C₁-C₃ alkyl. Nitration ofhalo-substituted isoquinoline A is carried out using nitric acid in thepresence of sulfuric acid under reduced temperatures, e.g., 0° C. toproduce nitro-substituted compound B. Boronic acid R³B(OH)₂ is coupledto intermediate B via a Suzuki reaction using an appropriate palladiumcatalyst, e.g., Pd(dddf)Cl₂, in the presence of a suitable base, e.g.,CsCO₃ in dioxane, to generate R³-substituted compound C.Nitro-containing compound C is reduced to the amine D. Amine D isreacted with sodium nitrite under acidic conditions and then treatedwith CuCl₂ and SO₂ to give sulfonyl chloride E. The addition ofR¹-substituted aza-containing heteroaryl X typically proceeds underbasic conditions using an appropriate base such as TEA in a solvent suchas DCM to afford compounds F. Reaction with m-CPBA gives N-oxide G.Treating N-oxide G with acetic anhydride with heating gives the acetatethat undergoes spontaneous 2,3-sigmatropic rearrangement. Deprotectionof the acetate provides 1-oxo compound H.

Scheme III illustrates the preparation of compounds of formula (II) ofthe present invention, wherein R³ is halogen. 4-Haloisoquinoline A isnitrated, for example with with H₂SO₄ and KNO₃ to furnish nitrohaloisoquinoline compound B. Nitroisoquinoline B is treated withoxidant, for example m-CPBA to provide N-oxide C which is furtherreacted with a chlorinating agent such as POCl₃ to afforddihaloisoquinoline compound D. Compound D undergoes SNAr substitutionwith an oxygen nucleophile for example sodium methoxide to givemethoxyisoquinoline compound E. Compound E was reduced, for example withZn to isoquinolinamine compound F. Compound F was subjected to Sanmeyerreaction conditions for example with NaNO₂, HCl, SO₂ and CuCl to impartisoquinolinesulfonyl chloride compound G. The addition of R¹-substitutedaza-containing heteroaryl X to compound G typically proceeds under basicconditions for example pyridine in a solvent such as DCM to affordsulfonamide compound H. Compound H was deprotected for example withtrimethylsilyl iodide to cede isoquinolinone compound I of Formula II.

The following Examples are intended to illustrate further certainembodiments of the invention and are not intended to limit the scope ofthe invention.

Example 1

5-((4-Methylindolin-1-yl)sulfonyl)isoquinolin-1(2H)-one

Compound 1-2: A solution of 1-chloroisoquinoline (5 g, 30.5 mmol) insulfurochloridic acid (50 mL, 750.9 mmol) was stirred at 150° C. for 12h under an N₂ atmosphere. The reaction mixture was quenched by addingice and then was diluted with H₂O (300 mL) and extracted with DCM (100mL×3). The combined organic layers were washed with saturated brine (300mL), dried over anhydrous Na₂SO₄, filtered and concentrated underreduced pressure to give a residue. The residue was purified by columnchromatography (SiO₂, petroleum ether:ethyl acetate=0:1 to 10:1).Compound 1-2, 1-chloroisoquinoline-5-sulfonyl chloride (19.1 g, 72.9mmol, 47.7% yield), was obtained as a yellow solid.

Compound 1-3: To a solution of 1-chloroisoquinoline-5-sulfonyl chloride(2.5 g, 9.54 mmol) in HCl aq. solution (30 mL, 9 N, 276.9 mmol) wasstirred at 100° C. for 12 h under an N₂ atmosphere. The reaction mixturewas filtered and concentrated under reduced pressure to give a residue.The crude product, Compound 1-3, 1-hydroxyisoquinoline-5-sulfonic acid(2.8 g, crude), was obtained as a yellow solid and was used in the nextstep without further purification.

Compound 1-4: To a solution of 1-hydroxyisoquinoline-5-sulfonic acid(0.8 g, 3.55 mmol) in SOCl₂ (40 mL) was added DMF (273 μL, 3.55 mmol) at0° C. and the mixture was stirred at 15° C. for 30 min under an N₂atmosphere. Then the mixture was stirred at 35° C. for 2.5 h under an N₂atmosphere. The reaction mixture was diluted with H₂O (100 mL) andextracted with DCM (200 mL×3). The combined organic layers were washedwith saturated brine (100 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue. The crudeproduct, Compound 1-4, 1-hydroxyisoquinoline-5-sulfonyl chloride (0.3 g,1.23 mmol, 34.6% yield), was obtained as a yellow solid. The compoundwas used in the next step without further purification.

EXAMPLE 1: To a solution of 1-hydroxyisoquinoline-5-sulfonyl chloride(150 mg, 615.6 μmol) in DCM (10 mL) was added TEA (128.5 μL, 923.4 μmol)and 4-methylindoline (90.1 mg, 677.1 μmol). The mixture was stirred at15° C. for 5 h under an N₂ atmosphere. The reaction mixture was dilutedwith H₂O (60 mL) and extracted with DCM (200 mL×2). The combined organiclayers were washed with saturated brine (80 mL), dried over anhydrousNa₂SO₄, filtered and concentrated under reduced pressure to give aresidue. The residue was purified by column chromatography (SiO₂,petroleum ether:ethyl acetate=10:1 to 1:1). EXAMPLE 1,5-(4-methylindolin-1-yl)sulfonyl-2H-isoquinolin-1-one (90 mg, 252 μmol,41.1% yield), was obtained as a white solid. LC-MS: [M+1] 340.09.

¹H NMR (400 MHz, DMSO-d₆): δ 11.65 (br, 1H), 8.48 (d, J=8 Hz, 1H), 8.26(d, J=7.6 Hz, 1H), 7.59 (t, J=7.6 Hz, 1H), 7.34 (t, J=6 Hz, 1H),7.20˜7.02 (m, 3H), 6.81 (d, J=7.6 Hz, 1H), 4.04 (t, J=8.0 Hz, 2H), 2.89(t, J=8 Hz, 2H), 2.11 (s, 3H).

Example 2

5-((5-Hydroxyindolin-1-yl)sulfonyl)isoquinolin-1(2H)-one

Compound 2-2: To a solution of 1-chloroisoquinoline-5-sulfonyl chloride(0.4 g, 1.53 mmol) in DCM (10 mL) was added TEA (318 μL, 2.29 mmol) andindolin-5-ol (226.9 mg, 1.68 mmol). The mixture was stirred at 25° C.for 12 h under an N₂ atmosphere. The reaction mixture was filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by column chromatography (SiO₂, petroleum ether:ethylacetate=10:1 to 1:1). Compound 2-2, 1-[(1-chloro-5-isoquinolyl)sulfonyl]indolin-5-ol (350 mg, 970.0 μmol, 63.6% yield), was obtained asa yellow solid.

EXAMPLE 2: A solution of1-[(1-chloro-5-isoquinolyl)sulfonyl]indolin-5-ol (330 mg, 914.6 μmol) inHCl (10 mL, 9 N) was stirred at 90° C. for 12 h under an N₂ atmosphere.The reaction mixture was quenched by addition of saturated NaHCO₃solution until pH=7, and then diluted with H₂O (30 mL) and extractedwith EtOAc (10 mL×3). The combined organic layers were washed withsaturated brine (30 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by prep-HPLC (column: Waters Xbridge Prep OBD C18 150 mm×40mm×10 m; mobile phase: [A, water (10 mM NH₄HCO₃)—B, ACN]; B %: 25%-55%,11 min). EXAMPLE 2,5-(5-hydroxyindolin-1-yl)sulfonyl-2H-isoquinolin-1-one (6 mg, 17.1 μmol,1.87% yield), was obtained as a yellow solid. LC-MS: [M+1] 342.07.

¹H NMR (400 MHz, CDCl3): δ 10.16 (br, 1H), 8.73 (d, J=8 Hz, 1H), 8.20(d, J=8.8 Hz, 1H), 7.56˜7.44 (m, 2H), 7.38˜7.30 (m, 1H), 6.74 (s, 1H),6.43 (d, J=6.4 Hz, 1H), 6.35 (d, J=8.4 Hz, 1H), 3.54 (t, J=8.4 Hz, 2H),2.93 (d, J=8.4 Hz, 2H). One proton was exchanged.

Example 3

5-(Indolin-1-ylsulfonyl)isoquinolin-1(2H)-one

Compound 3-1: To a solution of 1-chloroisoquinoline-5-sulfonyl chloride(0.3 g, 1.14 mmol) in DCM (8 mL) was added TEA (238.9 μL, 1.72 mmol) andindoline (109.1 mg, 915.6 tmol). The mixture was stirred at 15° C. for12 h under an N₂ atmosphere. The reaction mixture was diluted with H₂O(60 mL) and extracted with EtOAc (100 mL×3). The combined organic layerswere washed with saturated brine (50 mL), dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to give a residue. Thecrude product, Compound 3-1,1-chloro-5-indolin-1-ylsulfonyl-isoquinoline (0.2 g, crude) was obtainedas a yellow solid. Compound 3-1 was used in the next step withoutfurther purification.

EXAMPLE 3: A mixture of 1-chloro-5-indolin-1-ylsulfonyl-isoquinoline(0.2 g, 580.02 μmol, 1 eq) in HCl aq. solution (5 mL, 12M) was stirredat 100° C. for 12 h under an N₂ atmosphere. The reaction mixture wasquenched by addition of ice water (15 mL) at 0° C., and then NaHCO₃ wasadded to the mixture to adjust to pH 8 and filtered. The filtrate wasconcentrated under reduced pressure to give a residue. The residue waspartitioned between H₂O (60 mL) and DCM (100 mL×3). The combined organiclayers were washed with saturated brine (50 mL), dried over anhydrousNa₂SO₄, filtered and concentrated under reduced pressure to give aresidue. The residue was purified by column chromatography (SiO₂,petroleum ether:ethyl acetate=10:1 to 0:1). EXAMPLE 3,5-indolin-1-ylsulfonyl-2H-isoquinolin-1-one (40 mg, 117.5 μmol, 20.2%yield), was obtained as a white solid. LC-MS: [M+1] 326.07.

¹H NMR (400 MHz, MeOD): δ 8.58 (d, J=8.4 Hz, 1H), 8.34 (d, J=8 Hz, 1H),7.61 (t, J=8 Hz, 1H), 7.47 (d, J=8 Hz, 1H), 7.38 (d, J=7.6 Hz, 1H),7.25˜7.10 (m, 3H), 7.00 (t, J=8 Hz, 1H), 4.03 (t, J=8 Hz, 2H), 2.85 (t,J=8.4 Hz, 2H). One proton was exchanged.

Example 4

5-((6-Chloroindolin-1-yl)sulfonyl)isoquinolin-1(2H)-one

EXAMPLE 4: To a solution of 1-hydroxyisoquinoline-5-sulfonyl chloride(100.0 mg, 410.4 μmol) in DCM (5 mL) was added TEA (85.7 μL, 615.6 μmol)and 6-chloroindoline Indoline 2 (56.7 mg, 369.4 μmol). The mixture wasstirred at 15° C. for 12 h under an N₂ atmosphere. The reaction mixturewas diluted with H₂O (50 mL) and extracted with DCM (150 mL×2). Thecombined organic layers were washed with saturated brine (100 mL), driedover anhydrous Na₂SO₄, filtered and concentrated under reduced pressureto give a residue. The residue was purified by column chromatography(SiO₂, Pether:ethyl acetate=8:1 to 0:1). 40 mg of EXAMPLE 4,5-(6-chloroindolin-1-yl) sulfonyl-2H-isoquinolin-1-one, was obtained asa white solid. LC-MS: [M+1] 360.03.

¹H NMR (400 MHz, MeOD): δ 8.60 (d, J=8 Hz, 1H), 8.35 (d, J=8 Hz, 1H),7.63 (t, J=8 Hz, 1H), 7.45 (d, J=2 Hz, 1H), 7.37 (d, J=7.6 Hz, 1H), 7.26(d, J=7.6 Hz, 1H), 7.10 (d, J=7.6 Hz, 1H), 7.02˜6.96 (m, 1H), 4.03 (t,J=8.4 Hz, 2H), 2.87 (t, J=8.4 Hz, 2H). One proton was exchanged.

Example 5

5-((6-Fluoroindolin-1-yl)sulfonyl)isoquinolin-1(2H)-one

EXAMPLE 5: To a solution of 1-hydroxyisoquinoline-5-sulfonyl chloride(150.0 mg, 615.6 μmol) in DCM (5 mL) was added 6-fluoroindoline 5A (75.9mg, 554.0 μmol) and TEA (128.5 μL, 923.3 μmol). The mixture was stirredat 15° C. for 12 h under an N₂ atmosphere. The reaction mixture wasdiluted with H₂O (30 mL) and extracted with DCM (100 mL×2). The combinedorganic layers were washed with saturated brine (50 mL), dried overanhydrous Na₂SO₄, filtered and concentrated under reduced pressure togive a residue. The residue was purified by column chromatography (SiO₂,petroleum ether:ethyl acetate=10:1 to 0:1). EXAMPLE 5,5-(6-fluoroindolin-1-yl)sulfonyl-2H-isoquinolin-1-one (80 mg, 204.3μmol, 33.2% yield), was obtained as a yellow solid. LC-MS: [M+1] 344.06.

¹H NMR (400 MHz, MeOD): δ 8.59 (d, J=8.0 Hz, 1H), 8.36 (d, J=7.6 Hz,1H), 7.62 (t, J=8 Hz, 1H), 7.38 (d, J=8 Hz, 1H), 7.26 (d, J=7.6 Hz, 1H),7.22˜7.17 (m, 1H), 7.12˜7.06 (m, 1H), 6.75˜6.66 (m, 1H), 4.03 (t, J=8.4Hz, 2H), 2.87 (t, J=8.4 Hz, 2H). One proton was exchanged.

Example 6

5-((6-Methylindolin-1-yl)sulfonyl)isoquinolin-1(2H)-one

Compound 6-1: To a solution of 1-chloroisoquinoline-5-sulfonyl chloride(0.3 g, 1.14 mmol) in DCM (8 mL) was added TEA (239 μL, 1.72 mmol) and6-methylindoline 6A (121 mg, 916 μmol). The mixture was stirred at 15°C. for 12 h under an N₂ atmosphere. The reaction mixture was dilutedwith H₂O (60 mL) and extracted with EtOAc (100 mL×3). The combinedorganic layers were washed with saturated brine (50 mL), dried overanhydrous Na₂SO₄, filtered and concentrated under reduced pressure togive a residue. The crude product, compound 6-1,1-chloro-5-(6-methylindolin-1-yl)sulfonyl-isoquinoline (0.2 g, crude),was obtained as a yellow solid. The crude product was used in the nextstep without further purification.

EXAMPLE 6: A mixture of 1-chloro-5-(6-methylindolin-1-yl)sulfonyl-isoquinoline (0.2 g, 557 μmol) in HCl aq. solution (5 mL, 12 M)was degassed and purged with N₂ 3 times, and then the mixture wasstirred at 100° C. for 12 h under an N₂ atmosphere. The reaction mixturewas quenched by the addition of ice water (15 mL) at 0° C., and thenNaHCO₃ was added to the mixture to adjust to pH 8 and filtered. Thefiltrate was concentrated under reduced pressure to give a residue. Thereaction mixture was diluted with H₂O (60 mL) and extracted with DCM(100 mL×3). The combined organic layers were washed with saturated brine(50 mL), dried over anhydrous Na₂SO₄, filtered and concentrated underreduced pressure to give a residue. The residue was purified by columnchromatography (SiO₂, petroleum ether:ethyl acetate=10:1 to 0:1).EXAMPLE 6, 5-(6-methylindolin-1-yl)sulfonyl-2H-isoquinolin-1-one (40 mg,107.4 μmol, 19.2% yield), was obtained as a white solid. LC-MS: [M+1]340.09.

¹H NMR (400 MHz, MeOD): δ 8.57 (d, J=8.4 Hz, 1H), 8.32 (d, J=7.6 Hz,1H), 7.59 (t, J=8 Hz, 1H), 7.38 (d, J=8 Hz, 1H), 7.30 (s, 1H), 7.22 (d,J=8 Hz, 1H), 6.99 (d, J=7.6 Hz, 1H), 6.82 (d, J=7.6 Hz, 1H), 4.01 (t,J=8 Hz 2H), 2.79 (t, J=8 Hz 2H), 2.31 (s, 3H). One proton was exchanged.

Example 7

5-((4-(Hydroxymethyl)indolin-1-yl)sulfonyl)isoquinolin-1(2H)-one

EXAMPLE 7: To a solution of 1-oxo-2H-isoquinoline-5-sulfonyl chloride(0.3 g, 1.23 mmol) in DCM (10 mL) was added TEA (257.0 μL, 1.85 mmol)and indolin-4-ylmethanol 7A (202.0 mg, 1.35 mmol). The mixture wasstirred at 25° C. for 12 h under an N₂ atmosphere. The reaction mixturewas diluted with H₂O (15 mL) and extracted with DCM (5 mL×3). Thecombined organic layers were washed with saturated brine (15 mL), driedover anhyrdous Na₂SO₄, filtered and concentrated under reduced pressureto give a residue. The residue was purified by column chromatography(SiO₂, petroleum ether:ethyl acetate=1:0 to 0:1). EXAMPLE 7,5-[4-(hydroxymethyl)indolin-1-yl] sulfonyl-2H-isoqu-inolin-1-one (35 mg,91.59 mol, 7.44% yield), was obtained as a black brown solid. LC-MS:[M+1] 356.08.

¹H NMR (400 MHz, MeOD): δ 8.56 (d, J=8.0 Hz, 1H), 8.34 (d, J=7.6 Hz,1H), 7.58 (t, J=7.6 Hz, 1H), 7.50 (d, J=8 Hz, 2H), 7.22 (d, J=8 Hz, 1H),7.17 (t, J=8 Hz, 1H), 7.04 (d, J=7.6 Hz, 1H), 4.46 (s, 2H), 4.06 (t,J=8.4 Hz, 2H), 2.91 (t, J=8.4 Hz, 2H). Two protons were exchanged.

Example 8

5-((4-(Trifluoromethyl)indolin-1-yl)sulfonyl)isoquinolin-1(2H)-one

EXAMPLE 8: To a solution of 1-hydroxyisoquinoline-5-sulfonyl chloride(150 mg, 615 μmol) in DCM (5 mL) was added TEA (128.5 μL, 923.3 μmol)and 4-(trifluoromethyl)indoline 8A (103.6 mg, 554.0 μmol). The mixturewas stirred at 15° C. for 12 h under an N₂ atmosphere. The reactionmixture was diluted with H₂O (30 mL) and extracted with DCM (75 mL×2).The combined organic layers were washed with saturated brine (20 mL),dried over anhydrous Na₂SO₄, filtered and concentrated under reducedpressure to give a residue. The residue was purified by columnchromatography (SiO₂, petroleum ether:ethyl acetate=10:1 to 0:1).EXAMPLE 8,5-[4-(trifluoromethyl)indolin-1-yl]sulfonyl-2H-isoquinolin-1-one (20 mg,44.2 μmol, 7.18% yield), was obtained as a white solid. LC-MS: [M+1]394.06.

¹H NMR (400 MHz, MeOD): δ 8.60 (d, J=8 Hz, 1H), 8.38 (d, J=7.6 Hz, 1H),7.70 (d, J=8.4 Hz, 1H), 7.63 (t, J=8 Hz, 1H), 7.40˜7.20 (m, 4H), 4.10(t, J=8.4 Hz, 2H), 3.05 (t, J=8.4 Hz, 2H). One proton was exchanged.

Example 9

5-((4-Cyclopropylindolin-1-yl)sulfonyl)isoquinolin-1(2H)-one

EXAMPLE 9: To a solution of 1-hydroxyisoquinoline-5-sulfonyl chloride(150 mg, 615.5 μmol) in DCM (5 mL) was added TEA (128.5 μL, 923.3 μmol)and 4-cyclopropylindoline INDOLINE 22 (88.2 mg, 554.0 μmol). The mixturewas stirred at 15° C. for 12 h under an N₂ atmosphere. The reactionmixture was diluted with H₂O (40 mL) and was extracted with DCM (100mL×2). The combined organic layers were washed with saturated brine (20mL), dried over anhydrous Na₂SO₄, filtered and concentrated underreduced pressure to give a residue. The residue was purified by columnchromatography (SiO₂,petroleum ether:ethyl acetate=10:1 to0:1). EXAMPLE9, 5-(4-cyclopropylindolin-1-yl)sulfonyl-2H-isoqui-nolin-1-one (25 mg,57.8 μmol, 9.39% yield), was obtained as a white solid. LC-MS: [M+1]366.10.

¹H NMR (400 MHz, MeOD): δ 8.57 (d, J=7.6 Hz, 1H), 8.33 (d, J=7.6 Hz,1H), 7.59 (t, J=8 Hz, 1H), 7.33 (d, J=8 Hz, 1H), 7.26 (d, J=8 Hz, 1H),7.19 (d, J=8 Hz, 1H), 7.07 (t, J=7.6 Hz, 1H), 6.56 (d, J=7.6 Hz, 1H),4.05 (t, J=8.4 Hz, 1H), 2.93 (t, J=8.4 Hz, 2H), 1.73˜1.67 (m, 1H),0.88˜0.86 (m, 2H), 0.57˜0.55 (m, 2H).

Example 10

5-((4-ethylindolin-1-yl)sulfonyl)isoquinolin-1(2H)-one

Compound 10-1: To a solution of 1-hydroxyisoquinoline-5-sulfonylchloride (150 mg, 615.5 μmol) in DCM (5 mL) was added TEA (128.5 μL,923.3 μmol) and 4-vinylindoline INDOLINE 20 (98.3 mg, 677.1 μmol). Themixture was stirred at 15° C. for 12 h under an N₂ atmosphere. Thereaction mixture was diluted with H₂O (20 mL) and extracted with DCM (80mL×2). The combined organic layers were washed with saturated brine (30mL), dried over anhydrous Na₂SO₄, filtered and concentrated underreduced pressure to give a residue. The residue was purified by columnchromatography (SiO₂, petroleum ether:ethyl acetate=10:1 to 0:1).Compound 10-1, 5-(4-vinylindolin-1-yl)sulfonyl-2H-isoquinolin-1-one (50mg, 141.8 μmol, 23.05% yield), was obtained as a yellow solid.

EXAMPLE 10: A mixture of5-(4-vinylindolin-1-yl)sulfonyl-2H-isoquinolin-1-one (50 mg, 141.8μmol), Pd/C (5 mg, 10 wt %) in MeOH (5 mL) was degassed and purged withH₂ 3 times, and then the mixture was stirred at 15° C. for 12 h under H₂atmosphere (15 psi). The reaction mixture was filtered and the filterwas concentrated. The residue was purified by column chromatography(SiO₂, petroleum ether:ethyl acetate=5:1 to 0:1). The residue waspurified by prep-HPLC (column: Waters Xbridge 150 mm×50 mm×10 μm; mobilephase:[A, water (10 mM NH₄HCO₃)—B, ACN]; B %: 30%-60%, 12 min). EXAMPLE10, 5-(4-ethylindolin-1-yl)sulfonyl-2H-isoquinolin-1-one (14 mg, 39.4μmol, 27.7% yield), was obtained as a white solid. LC-MS: [M+1] 354.10.

¹H NMR (400 MHz, MeOD): δ 8.58 (d, J=7.6 Hz, 1H), 8.34 (d, J=7.6 Hz,1H), 7.60 (t, J=8 Hz, 1H), 7.32 (t, J=7.2 Hz, 2H), 7.18 (d, J=4.0 Hz,1H), 7.13 (t, J=7.6 Hz, 1H), 6.88 (d, J=4 Hz, 1H), 4.03 (t, J=8.4 Hz,2H), 2.76 (t, J=8.4 Hz, 2H), 2.50˜2.44 (m, 2H), 1.06 (t, J=7.6 Hz, 3H).One proton was exchanged.

Example 11

5-((4-(1-Hydroxyethyl)indolin-1-yl)sulfonyl)isoquinolin-1(2H)-one

EXAMPLE 11: To a solution of 1-oxo-2H-isoquinoline-5-sulfonyl chloride(244.2 mg, 1.00 mmol) in DCM (5 mL) was added TEA (209.3 μL, 1.50 mmol)and 1-indolin-4-ylethanol INDOLINE 8 (180 mg, 1.10 mmol). The mixturewas stirred at 25° C. for 12 h under an N₂ atmosphere. The reactionmixture was diluted with H₂O (60 mL) and extracted with DCM (150 mL×2).The combined organic layers were washed with saturated brine (50 mL),dried over anhydrous Na₂SO₄, filtered and concentrated under reducedpressure to give a residue. The residue was purified by columnchromatography (SiO₂, petroleum ether:ethyl acetate=10:1 to 0:1). Theresidue was purified by prep-HPLC (column: Waters Xbridge 150 mm×25 mm×5μm; mobilephase:[A, water (10 mM NH₄HCO₃)—B, ACN]; B %: 10%-40%, 10min). EXAMPLE 11,5-[4-(1-hydroxyethyl)indolin-1-yl]sulfonyl-2H-isoquinolin-1-one (40 mg),was obtained as a white solid. LC-MS: [M+1] 370.10.

¹H NMR (400 MHz, MeOD): δ 8.57 (d, J=7.6 Hz, 1H), 8.35 (d, J=8 Hz, 1H),7.59 (t, J=8 Hz, 1H), 7.38 (d, J=8, 1H), 7.34 (d, J=8 Hz, 1H), 7.21-7.15(m, 2H), 7.11 (d, J=7.6 Hz, 1H), 4.88-4.71 (m, 1H), 4.07-4.01 (m, 2H),2.89-2.80 (m, 2H), 1.26 (d, J=6.4 Hz, 3H). Two protons were exchanged.

Example 12

5-((3,3-Dimethylindolin-1-yl)sulfonyl)isoquinolin-1(2H)-one

Compound 12-1: To a solution of 1-chloroisoquinoline-5-sulfonyl chloride(0.3 g, 1.14 mmol) in DCM (8 mL) was added TEA (238.9 μL, 1.72 mmol) and3,3-dimethylindoline 12A (134.8 mg, 915.6 μmol). The mixture was stirredat 15° C. for 12 h under an N₂ atmosphere. The reaction mixture wasdiluted with H₂O (60 mL) and extracted with EtOAc (100 mL×3). Thecombined organic layers were washed with saturated brine (50 mL), driedover anhydrous Na₂SO₄, filtered and concentrated under reduced pressureto give a residue. The crude product Compound 12-1,1-chloro-5-(3,3-dimethylindolin-1-yl)sulfonyl-isoquinoline (0.2 g,crude), was obtained as a yellow solid. The crude product was used inthe next step without further purification.

EXAMPLE 12: A mixture of1-chloro-5-(3,3-dimethylindolin-1-yl)sulfonyl-isoquinoline (0.2 g, 536.3μmol) in HCl aq. solution (5 mL, 12 M) was degassed and purged with N₂ 3times, and then the mixture was stirred at 100° C. for 12 h under an N₂atmosphere. The reaction mixture was quenched by the addition of icewater (15 mL) at 0° C., and then NaHCO₃ was added to the mixture toadjust to pH 8, filtered and the filtrate was concentrated under reducedpressure to give a residue. The residue was diluted with H₂O (60 mL) andextracted with DCM (100 mL×3). The combined organic layers were washedwith saturated brine (50 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by column chromatography (SiO₂, petroleum ether:ethylacetate=10:1 to 0:1). EXAMPLE 12,5-(3,3-dimethylindolin-1-yl)sulfonyl-2H-isoquinolin-1-one (12 mg, 31.5μmol, 5.88% yield), was obtained as a white solid. LC-MS: [M+1] 354.10.

¹H NMR (400 MHz, MeOD): δ 8.58, (d, J=8 Hz, 1H), 8.42 (d, J=8 Hz, 1H),7.62 (t, J=7.6 Hz, 1H), 7.48 (d, J=8 Hz, 1H), 7.43 (d, J=7.6 Hz, 1H),7.23 (d, J=7.6 Hz, 1H), 7.18 (t, J=7.6 Hz, 1H), 7.11 (d, J=7.6 Hz, 1H),7.02 (t, J=7.6 Hz, 1H), 3.73 (s, 2H), 1.08 (s, 6H). One proton wasexchanged.

Example 13

5-((3-Methylindolin-1-yl)sulfonyl)isoquinolin-1(2H)-one

EXAMPLE 13: To a solution of 1-hydroxyisoquinoline-5-sulfonyl chloride(100 mg, 410.4 μmol) in DCM (5 mL) was added TEA (85.6 μL, 615.6 μmol)and 3-methylindoline (55.7 mg, 328.3 μmol). The mixture was stirred at15° C. for 12 h under an N₂ atmosphere. The reaction mixture was dilutedwith H₂O (50 mL) and extracted with DCM (150 mL×2). The combined organiclayers were washed with saturated brine (50 mL), dried over anhydrousNa₂SO₄, filtered and concentrated under reduced pressure to give aresidue. The residue was purified by column chromatography(SiO₂,petroleum ether:ethyl acetate=10:1 to 0:1). 70 mg of EXAMPLE 13,5-(3-methylindolin-1-yl) sulfony 1-2H-isoquinolin-1-one, was obtained asa white solid. LC-MS: [M+1] 340.09.

¹H NMR (400 MHz, DMSO-d₆): δ 11.67 (s, 1H), 8.49 (d, J=8 Hz, 1H), 8.30(d, J=8 Hz, 1H), 7.61 (t, J=7.6 Hz, 1H), 7.31˜7.30 (m, 2H), 7.20˜7.13(m, 3H), 7.01 (t, J=7.6 Hz, 1H), 4.19 (t, J=9.2 Hz, 1H), 3.57-3.53 (m,1H), 3.29˜3.24 (m, 1H), 1.05 (d, J=6.8 Hz, 3H).

Example 14

5-((4-Isopropylindolin-1-yl)sulfonyl)isoquinolin-1(2H)-one

Compound 14-1: To a solution of 1-hydroxyisoquinoline-5-sulfonylchloride (83.4 mg, 342.6 μmol) in DCM (5 mL) was added TEA (71.5 μL,513.8 μmol) and 4-isopropenylindoline INDOLINE 18 (60 mg, 376.82 μmol).The mixture was stirred at 15° C. for 12 h under an N₂ atmosphere. Thereaction mixture was diluted with H₂O (10 mL) and extracted with DCM (50mL×3). The combined organic layers were washed with saturated brine (10mL), dried over anhydrous Na₂SO₄, filtered and concentrated underreduced pressure to give a residue. The residue was purified by columnchromatography (SiO₂, petroleum ether:ethyl acetate=10:1 to 1:1).Compound 14-1,5-(4-isopropenylindolin-1-yl)sulfonyl-2H-isoquinolin-1-one(50 mg, 136.4 μmol, 39.8% yield), was obtained as a yellow solid.

EXAMPLE 14: To a solution of5-(4-isopropenylindolin-1-yl)sulfonyl-2H-isoquinolin-1-one (50 mg,136.45 μmol, 1 eq) in MeOH (5 mL) was added Pd/C (5 mg, 10% purity)under H₂. The mixture was stirred at 15° C. for 12 h with a pressure of15 psi. The reaction mixture was filtered and the filtrate wasconcentrated. The residue was purified by column chromatography (SiO₂,petroleum ether:ethyl acetate=5:1 to 0:1). The residue was purified byprep-HPLC (column: Waters Xbridge 150 mm×50 mm×10 um; mobile phase: [A,water (10 mM NH₄HCO₃)—B, ACN];B %: 30%-60%, 12 min). EXAMPLE 14,5-(4-isopropylindolin-1-yl)sulfonyl-2H-isoquinolin-1-one (8 mg, 21.50μmol, 15.7% yield), was obtained as a white solid. LC-MS: [M+1] 368.12.

¹H NMR (400 MHz, MeOD): δ 8.57 (d, J=8.0 Hz, 1H), 8.34 (d, J=6.8 Hz,1H), 7.59 (t, J=8 Hz, 1H), 7.32-7.26 (m, 2H), 7.15 (t, J=7.6 Hz, 2H),6.95 (d, J=7.6 Hz, 1H), 4.00 (t, J=8 Hz, 2H), 2.81˜2.71 (m, 3H), 1.10(d, J=6.8 Hz, 6H). One proton was exchanged.

Example 15

1-((1-Oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile

EXAMPLE 15: To a solution of 1-hydroxyisoquinoline-5-sulfonyl chloride(202.8 mg, 832.3 μmol) in DCM (5 mL) was added TEA (173.8 μL, 1.25 mmol)and indoline-6-carbonitrile INDOLINE 4 (120 mg, 832.3 μmol). The mixturewas stirred at 25° C. for 5 h under an N₂ atmosphere. The reactionmixture was quenched by addition H₂O (40 mL), then the reaction mixturewas extracted with DCM (100 mL×2). The combined organic layers werewashed with saturated brine 50 (mL), dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to give a residue. Theresidue was purified by column chromatography (SiO₂, petroleumether:ethyl acetate=10:1 to 0:1). EXAMPLE 15,1-[(1-oxo-2H-isoquinolin-5-yl)sulfonyl]indoline-6-carbonitrile (80 mg,215.8 μmol, 25.9% yield), was obtained as a yellow solid. LC-MS: [M+1]351.07.

¹H NMR (400 MHz, DMSO-d₆): δ 11.68 (s, 1H), 8.51 (d, J=7.6 Hz, 1H), 8.37(d, J=6.4 Hz, 1H), 7.64 (t, J=8 Hz, 1H), 7.58 (s, 1H), 7.47-7.40 (m,3H), 7.11 (d, J=7.2 Hz, 1H), 4.06 (t, J=8 Hz, 2H), 3.07 (t, J=8.0 Hz,2H).

Example 16

5-((4,6-Dichloroindolin-1-yl)sulfonyl)isoquinolin-1(2H)-one

EXAMPLE 16: To a solution of 1-oxo-2H-isoquinoline-5-sulfonyl chloride(176.6 mg, 725.1 μmol) in DCM (3 mL) was added TEA (151.3 μL, 1.09 mmol)and 4,6-dichloroindoline INDOLINE 10 (150 mg, 797.6 μmol). The mixturewas stirred at 25° C. for 12 h under an N₂ atmosphere. The reactionmixture was diluted with H₂O (60 mL) and extracted with DCM (150 mL×2).The combined organic layers were washed with saturated brine (50 mL),dried over anhydrous Na₂SO₄, filtered and concentrated under reducedpressure to give a residue. The residue was purified by columnchromatography (SiO₂, petroleum ether:ethyl acetate=10:1 to 0:1). Theresidue was purified by prep-HPLC (column: Waters Xbridge Prep OBD C18150 mm×40 mm×10 μm; mobile phase: [A, water (10 mM NH₄HCO₃)—B,ACN];B %:35%-55%, 11 min). EXAMPLE 16,5-(4,6-dichloroindolin-1-yl)sulfonylisoquinolin-1-ol (60 mg, 150.5 μmol,20.7% yield), was obtained as a white solid. LC-MS: [M+1] 393.99.

¹H NMR (400 MHz, DMSO-d₆): δ 8.53 (d, J=7.6 Hz, 1H), 8.31 (d, J=8 Hz,2H), 7.67 (t, J=8 Hz, 1H), 7.39 (d, J=7.6 Hz, 1H) 7.23 (d, J=8.8 Hz,2H), 7.10 (d, J=7.6 Hz, 1H), 4.09 (t, J=8.4 Hz, 2H), 3.03 (t, J=8.4 Hz,2H).

Example 17

4-Cyclopropyl-1-((1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile

EXAMPLE 17: To a solution of 1-oxo-2H-isoquinoline-5-sulfonyl chloride(50 mg, 205.2 μmol) in DCM (5 mL) was added TEA (85.6 μL, 615.6 μmol)and 4-cyclopropylindoline-6-carbonitrile INDOLINE 13 (37.8 mg, 205.2μmol). The mixture was stirred at 25° C. for 12 h under an N₂atmosphere. The reaction mixture was diluted with H₂O (15 mL) andextracted with DCM (80 mL×2). The combined organic layers were washedwith saturated brine (10 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by prep-HPLC (column: Xtimate C18 150 mm×25 mm×5 m; mobilephase: [A, water (10 mM NH₄HCO₃)—B, ACN]; B %: 50%-80%, 10 min). EXAMPLE17,4-cyclopropyl-1-[(1-hydroxy-5-isoquinolyl)sulfonyl]indoline-6-carbonitrile(19 mg, 47.5 μmol, 23.1% yield), was obtained as a yellow solid. LC-MS:[M+1] 391.10.

¹H NMR (400 MHz, MeOD): δ 8.62 (d, J=8 Hz, 1H), 8.39 (d, J=6.8 Hz, 1H),7.65 (t, J=7.8 Hz, 1H), 7.54 (s, 1H), 7.35 (d, J=7.6 Hz, 1H), 7.27 (d,J=7.6 Hz, 1H), 6.92 (s, 1H), 4.09 (t, J=8.4 Hz, 2H), 3.07 (t, J=8.4 Hz,2H), 1.79˜1.77 (m, 1H), 0.98˜0.95 (m, 2H), 0.68-0.66 (m, 2H). One protonwas exchanged.

Example 18

4-Methyl-1-((1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile

EXAMPLE 18: To a solution of 1-oxo-2H-isoquinoline-5-sulfonyl chloride(100 mg, 410.4 μmol) in DCM (5 mL) was added TEA (65.9 μL, 474.0 μmol)and 4-methylindoline-6-carbonitrile INDOLINE 16 (50 mg, 316.0 μmol). Thereaction mixture was stirred at 25° C. for 12 h under an N₂ atmosphere.The reaction mixture was diluted with H₂O (20 mL) and was extracted withDCM (100 mL×2). The combined organic layers were washed with saturatedbrine (10 mL), dried over anhydrous Na₂SO₄, filtered and concentratedunder reduced pressure to give a residue. The residue was purified bycolumn chromatography (SiO₂, petroleum ether:ethyl acetate=1:0 to 0:1).The residue was purified by column chromatography (SiO₂, petroleumether:ethyl acetate=1:0 to 4:5). EXAMPLE 18,1-[(1-hydroxy-5-isoquinolyl)sulfonyl]-4-methyl-indoline-6-carbonitrile(8 mg, 19.9 μmol, 6.30% yield), was obtained as a white solid. LC-MS:[M+1] 365.08.

¹H NMR (400 MHz, MeOD): δ 8.61 (d, J=8 Hz, 1H), 8.38 (d, J=8 Hz, 1H),7.64 (t, J=8 Hz, 1H), 7.56 (s, 1H), 7.37 (d, J=7.6 Hz, 1H), 7.28 (d, J=8Hz, 1H), 7.21 (s, 1H), 4.07 (t, J=8.4 Hz, 2H), 2.96 (t, J=8.4 Hz, 2H),2.20 (s, 3H). One proton was exchanged.

Example 19

7-Chloro-5-((6-chloroindolin-1-yl)sulfonyl)isoquinolin-1(2H)-one

Compound 19-2: A mixture of 3,5-dichlorobenzaldehyde (25 g, 142.8 mmol),2,2-diethoxyethanamine (83.08 mL, 571.3 mmol) in toluene (200 mL) wasdegassed and purged with N₂ 3 times, and then the mixture was stirred at110° C. for 12 h under an N₂ atmosphere. The reaction mixture wasdiluted with H₂O (500 mL) and extracted with EtOAc (900 mL×2). Thecombined organic layers were washed with saturated brine (200 mL), driedover anhydrous Na₂SO₄, filtered and concentrated under reduced pressureto give a residue. The crude product, compound 19-2,(E)-1-(3,5-dichlorophenyl)-N-(2,2-diethoxyethyl)methanimine (35 g,crude) was obtained as a yellow solid. The crude product was used in thenext step without further purification.

Compound 19-3: To a solution of H₂SO₄ (500 mL) was added drop wise(E)-1-(3,5-dichlorophenyl)-N-(2,2-diethoxyethyl) methanimine (30 g,103.3 mmol) in DCM (100 mL) at 125° C. The resulting mixture was stirredat 125° C. for 6 h under an N₂ atmosphere. The reaction mixture wasquenched by addition of ice water (200 mL) at 0° C. and then aq.NaOH(200 mL) was added to the mixture to adjust pH 8. It was extracted withEtOAc (1000 mL×2). The combined organic layers were washed withsaturated brine (200 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by column chromatography (SiO₂, petroleum ether:ethylacetate=20:1 to 5:1). Compound 19-3, 5,7-dichloroisoquinoline (8 g, 40.3mmol, 39.0% yield), was obtained as a yellow solid.

Compound 19-4: A mixture of 5,7-dichloroisoquinoline (100 mg, 504.9μmol), diphenylmethanimine (137.2 mg, 757.3 μmol),(5-diphenylphosphanyl-9,9-dimethyl-xanthen-4-yl)-diphenyl-phosphane,methanesulfonate, [2-[2-(methylamino)phenyl]phenyl] palladium (485.9 mg,504.9 μmol), t-BuONa (2 M, 1.26 mL) in toluene (10 mL) was degassed andpurged with N₂ 3 times, and then the mixture was stirred at 100° C. for12 h under an N₂ atmosphere. The reaction mixture was diluted with H₂O(20 mL) and extracted with DCM (80 mL×2). The combined organic layerswere washed with saturated brine (10 mL), dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to give a residue. Theresidue was purified by column chromatography (SiO₂, petroleumether:ethyl acetate=20:1 to 1:1). Compound 19-4,N-(7-chloro-5-isoquinolyl)-1,1-diphenyl-methanimine (40 mg, 116.6 μmol,11.5% yield), was obtained as a yellow solid.

Compound 19-5: To a solution ofN-(7-chloro-5-isoquinolyl)-1,1-diphenyl-methanimine (0.3 g, 875.0 μmol)in THF (10 mL) was added HCl (3 M, 525.0 μL). The mixture was stirred at25° C. for 2 h under an N₂ atmosphere. The reaction mixture was quenchedby addition of ice water (10 mL) at 0° C., and then aq. NaOH (10 mL) wasadded to the mixture to adjust to pH 8. The reaction mixture was dilutedwith H₂O (10 mL) and extracted with DCM (50 mL×2). The combined organiclayers were washed with saturated brine (10 mL), dried over anhydrousNa₂SO₄, filtered and concentrated under reduced pressure to give aresidue. The residue was purified by column chromatography (SiO₂,petroleum ether:ethyl acetate=5:1 to 1:1). Compound 19-5,7-chloroisoquinolin-5-amine (0.1 g, 559.8 μmol, 63.9% yield), wasobtained as a yellow solid.

Compound 19-6: To a solution of 7-chloroisoquinolin-5-amine (0.1 g, 559μmol) in HCl (5 mL) was added NaNO₂ (38.6 mg, 559.8 μmol) and H₂O (1 mL)at −5° C. The mixture was stirred at −5° C. for 1 h. Then it wastransferred in one portion to a solution of AcOH (5 mL) with CuCl (13.8mg, 139.9 μmol) in H₂O (1 mL) that was saturated with bubbled SO₂. Themixture was stirred at 25° C. for 2 h. The reaction mixture wasconcentrated under reduced pressure to give a residue. The residue wastaken up with water (20 mL) and DCM and extracted with DCM (80 mL×2).The combined organic layers were washed with saturated brine (10 mL),dried over anhydrous Na₂SO₄, filtered and concentrated under reducedpressure to give a residue. The crude product, compound 19-6,7-chloroisoquinoline-5-sulfonyl chloride (0.2 g, crude) was obtained asa yellow solid that was used in the next step without furtherpurification.

Compound 19-7: To a solution of 7-chloroisoquinoline-5-sulfonyl chloride(0.2 g, 763.0 μmol) in acetone (10 mL) was added K₂CO₃ (158.1 mg, 1.14mmol) and 6-chloroindoline INDOLINE 2 (128.9 mg, 839.3 μmol). Themixture was stirred at 25° C. for 12 h under an N₂ atmosphere. Thereaction mixture diluted with H₂O (20 mL) and extracted with DCM (80mL×2). The combined organic layers were washed with saturated brine (10mL), dried over anhydrous Na₂SO₄, filtered and concentrated underreduced pressure to give a residue. The residue was purified by columnchromatography (SiO₂, petroleum ether:ethyl acetate=1:0 to 1:1).Compound 19-7, 7-chloro-5-(6-chloroindolin-1-yl)sulfonyl-isoquinoline(80 mg, 210.9 μmol, 27.6% yield), was obtained as a yellow solid.

Compound 19-8: A mixture of7-chloro-5-(6-chloroindolin-1-yl)sulfonyl-isoquinoline (80 mg, 210.9μmol), m-CPBA (65.7 mg, 316.4 μmol) in DCM (5 mL) was cooled to 0° C.,and then the mixture was stirred at 25° C. for 12 h under an N₂atmosphere. The reaction mixture was diluted with H₂O (15 mL) andextracted with DCM (25 mL×2). The combined organic layers were washedwith saturated brine 10 mL, dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by column chromatography (SiO₂, petroleum ether:EtOAc=10:1 toDCM:MeOH=10:1). Compound 19-8,7-chloro-5-(6-chloroindolin-1-yl)sulfonyl-2-oxido-isoquinolin-2-ium (50mg, 126.5 μmol, 59.9% yield), was obtained as a yellow solid.

Compound 19-9: A mixture of7-chloro-5-(6-chloroindolin-1-yl)sulfonyl-2-oxido-isoquinolin-2-ium (50mg, 126.5 μmol) in Ac₂O (3 mL) was warmed to 90° C., and then themixture was stirred at 90° C. for 3 h under an N₂ atmosphere. Thereaction mixture was concentrated under reduced pressure to give aresidue. The crude product[7-chloro-5-(6-chloroindolin-1-yl)sulfonyl-1-isoquinolyl] acetate (40mg, crude) was obtained as a brown solid that was used in the next stepwithout further purification.

EXAMPLE 19: A mixture of[7-chloro-5-(6-chloroindolin-1-yl)sulfonyl-1-isoquinolyl]acetate (40 mg,91.4 μmol) and KOH (7.70 mg, 137.2 μmol) in MeOH (5 mL) was degassed andpurged with N₂ 3 times, and then the mixture was stirred at 25° C. for 2h under an N₂ atmosphere. Then the mixture was stirred at 70° C. for 3 hunder an N₂ atmosphere. The reaction mixture was concentrated underreduced pressure to give a residue. The residue was purified byprep-HPLC (column: Xtimate C18 150 mm×25 mm×5 μm; mobile phase: [A,water (10 mM NH₄HCO₃)—B, ACN]; B %:50%-70%, 10 min). EXAMPLE 19,7-chloro-5-(6-chloroindolin-1-yl)sulfonyl-2H-isoquinolin-1-one (18 mg,45.0 μmol, 49.2% yield) was obtained as a yellow solid. LC-MS: [M+1]393.99.

¹H NMR (400 MHz, DMSO-d₆): δ 8.43 (d, J=4.4 Hz, 1H), 8.21 (d, J=2.8 Hz,1H), 7.38 (d, J=7.6 Hz, 1H), 7.33 (d, J=1.6 Hz, 1H), 7.23 (d, J=8 Hz,1H), 7.08 (t, J=7.6 Hz, 2H), 4.05 (t, J=8.4 Hz, 2H), 2.95 (t, J=8.4 Hz,2H). One proton was exchanged.

Example 20

1-((7-Chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile

Compound 20-1: To a solution of 7-chloroisoquinoline-5-sulfonyl chloride(0.1 g, 381.5 μmol) in acetone (8 mL) was added K₂CO₃ (79.0 mg, 572.2μmol) and indoline-6-carbonitrile INDOLINE 4 (60.5 mg, 419.6 μmol). Themixture was stirred at 25° C. for 12 h under an N₂ atmosphere. Thereaction mixture was diluted with H₂O (20 mL) and extracted with DCM (80mL×2). The combined organic layers were washed with saturated brine (10mL), dried over anhydrous Na₂SO₄, filtered and concentrated underreduced pressure to give a residue. The residue was purified by columnchromatography (SiO₂, petroleum ether:ethyl acetate=10:1 to 0:1).Compound 20-1,1-[(7-chloro-5-isoquinolyl)sulfonyl]indoline-6-carbonitrile (30 mg, 81.1μmol, 21.2% yield), was obtained as a yellow solid.

Compound 20-2: A mixture of1-[(7-chloro-5-isoquinolyl)sulfonyl]indoline-6-carbonitrile (30 mg, 81.1μmol), m-CPBA (26.2 mg, 121.6 μmol) in DCM (5 mL) was degassed andpurged with N₂ 3 times, and then the mixture was stirred at 25° C. for12 h under an N₂ atmosphere. The reaction mixture was diluted with H₂O(15 mL) and extracted with DCM (50 mL×2). The combined organic layerswere washed with saturated brine (10 mL), dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to give a residue. Theresidue was purified by column chromatography (SiO₂, petroleumether:ethyl acetate=5:1 to DCM:MeOH=10:1). Compound 20-2,1-(7-chloro-2-oxido-isoquinolin-2-ium-5-yl)sulfonylindoline-6-carbonitrile(30 mg), was obtained as a yellow solid.

Compound 20-3: A mixture of1-(7-chloro-2-oxido-isoquinolin-2-ium-5-yl)sulfonylindoline-6-carbonitrile(30 mg, 77.7 μmol) in Ac₂O (3 mL) was degassed and purged with N₂ 3times, and then the mixture was stirred at 100° C. for 12 h under an N₂atmosphere. The reaction mixture was concentrated under reduced pressureto give a residue. The crude product Compound 20-3,[7-chloro-5-(6-cyanoindolin-1-yl)sulfonyl-1-isoquinolyl] acetate (20 mg,crude), was obtained as a yellow oil that was used in the next stepwithout further purification.

EXAMPLE 20: A mixture of[7-chloro-5-(6-cyanoindolin-1-yl)sulfonyl-1-isoquinolyl]acetate (20 mg,46.7 μmol) and KOH (5.25 mg, 93.4 μmol) in MeOH (5 mL) was degassed andpurged with N₂ 3 times, and then the mixture was stirred at 25° C. for 2h under an N₂ atmosphere. The reaction mixture was diluted with H₂O (10mL) and extracted with DCM (50 mL×2). The combined organic layers werewashed with saturated brine (10 mL), dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to give a residue. Theresidue was purified by prep-HPLC (column: HUAPU C8 Extreme BDS 150mm×30 mm×5 um; mobile phase: [A, water (10 mM NH₄HCO₃)—B, ACN]; B %:45%-55%, 10 min). EXAMPLE 20,1-[(7-chloro-1-oxo-2H-isoquinolin-5-yl)sulfonyl]indoline-6-carbonitrile(2 mg, 4.7 μmol, 10.2% yield), was obtained as a white solid. LC-MS:[M+1] 385.03.

¹H NMR (400 MHz, DMSO-d₆): δ 8.44 (d, J=2.4 Hz, 1H), 8.32 (d, J=2 Hz,1H), 7.68 (s, 1H), 7.52 (d, J=1.2 Hz, 1H), 7.45˜7.36 (m, 2H), 7.04 (d,J=8.8 Hz, 1H), 4.07 (t, J=8.4 Hz, 2H), 3.07 (t, J=8.4 Hz, 2H). Oneproton was exchanged.

Example 21

5-((6-Chloroindolin-1-yl)sulfonyl)-4-fluoroisoquinolin-1-ol

Compound 21-2: To a solution of n-BuLi (2.5 M, 201.8 mL) in THF (100 mL)was added a solution of 4-bromoisoquinoline (30 g, 144.1 mmol) in THF(100 mL) at −65° C. The mixture was stirred at −65° C. for 30 min. Asolution of N-(benzenesulfonyl)-N-fluoro-benzenesulfonamide (100.0 g,317.2 mmol) in THF (100 mL) was added at −65° C. The mixture was stirredat −65° C. for 1 h. Then the mixture was stirred at 25° C. for 10.5 hunder an N₂ atmosphere. The reaction mixture was diluted with H₂O (500mL) and extracted with EtOAc (1500 mL×2). The combined organic layerswere washed with saturated brine (100 mL), dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to give a residue. Thecrude product, compound 21-2, 4-fluoroisoquinoline (30 g, crude), wasobtained as brown oil. The crude product was used in the next stepwithout further purification.

Compound 21-3: To a solution of 4-fluoroisoquinoline (8 g, 54.3 mmol) inH₂SO₄ (50 mL) was added KNO₃ (6.05 g, 59.8 mmol). The mixture wasstirred at 25° C. for 12 h under an N₂ atmosphere. The reaction mixturewas quenched by addition of ice water (100 mL) at −15° C., and thenaq.NaOH (50 mL) was added to the mixture to adjust to pH 8. It wasfiltered and the filtrate was concentrated under reduced pressure togive a residue. The residue was purified by column chromatography (SiO₂,petroleum ether:ethyl acetate=10:1 to 1:1). Compound 21-3,4-fluoro-5-nitro-isoquinoline (5 g, 26.0 mmol, 47.8% yield), wasobtained as a yellow solid.

Compound 21-4: A mixture of 4-fluoro-5-nitro-isoquinoline (5 g, 26.0mmol), NH₄Cl (11.1 g, 208.1 mmol), Zn (13.6 g, 208.1 mmol) in THF (20mL) and H₂O (10 mL) was degassed and purged with N₂ 3 times, and thenthe mixture was stirred at 25° C. for 5 h under an N₂ atmosphere. Thereaction mixture was filtered and the filtrate was concentrated. Theresidue was purified by column chromatography (SiO₂, petroleumether:ethyl acetate=10:1 to 1:1). Compound 21-4,4-fluoroisoquinolin-5-amine (3 g, 18.5 mmol, 71.1% yield), was obtainedas a yellow solid.

Compound 21-5: To a solution of 4-fluoroisoquinolin-5-amine (3 g, 18.5mmol) in HCl (10 mL) was added NaNO₃ (1.57 g, 18.5 mmol) and H₂O (2 mL)at −15° C. The mixture was stirred at −15° C. for 1 h under an N₂atmosphere. Then it was transferred to a solution of AcOH (10 mL) withCuCl (457.8 mg, 4.62 mmol) and H₂O (2 mL) that was saturated by bubbledSO₂. The mixture was stirred at −5° C. for 2 h. The reaction mixture wasconcentrated under reduced pressure to give a residue. The crude productCompound 21-5, 4-fluoroisoquinoline-5-sulfonyl chloride (1.1 g, crude),was obtained as a yellow solid and it was used in the next step withoutfurther purification.

Compound 21-6: A mixture of 4-fluoroisoquinoline-5-sulfonyl chloride(0.5 g, 2.04 mmol), 6-chloroindoline INDOLINE 2 (250.1 mg, 1.63 mmol),TEA (424.9 μL, 3.05 mmol) in DCM (10 mL) was degassed and purged with N₂3 times, and then the mixture was stirred at 25° C. for 12 h under an N₂atmosphere. The reaction mixture was diluted with H₂O (30 mL) and it wasextracted with DCM (100 mL×2). The combined organic layers were washedwith saturated brine (20 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by column chromatography (SiO₂, petroleum ether:ethylacetate=10:1 to 1:1). Compound 21-6,5-(6-chloroindolin-1-yl)sulfonyl-4-fluoro-isoquinoline (0.2 g, 551.2μmol), was obtained as a yellow solid.

Compound 21-7: A mixture of5-(6-chloroindolin-1-yl)sulfonyl-4-fluoro-isoquinoline (0.2 g, 551μmol), m-CPBA (142.6 mg, 826.9 μmol) in DCM (10 mL) was degassed andpurged with N₂ 3 times, and then the mixture was stirred at 25° C. for12 h under an N₂ atmosphere. The reaction mixture was diluted with H₂O(20 mL) and extracted with DCM (100 mL×2). The combined organic layerswere washed with saturated brine (10 mL), dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to give a residue. Theresidue was purified by column chromatography (SiO₂,petroleumether:ethyl acetate=5:1 to DCM:MeOH=10:1). Compound 21-7,5-(6-chloroindolin-1-yl)sulfonyl-4-fluoro-2-oxido-isoquinolin-2-ium (70mg, 184 μmol, 33.5% yield), was obtained as a white solid.

EXAMPLE 21: A mixture of5-(6-chloroindolin-1-yl)sulfonyl-4-fluoro-2-oxido-isoquinolin-2-ium (70mg, 184.7 μmol) in Ac₂O (10 mL) was degassed and purged with N₂ 3 times,and then the mixture was stirred at 110° C. for 8 h under an N₂atmosphere. The reaction mixture was diluted with H₂O (20 mL) andextracted with DCM (80 mL×2). The combined organic layers were washedwith saturated brine (10 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by column chromatography (SiO₂, petroleum ether:ethylacetate=10:1 to 0:1). The residue was purified by prep-HPLC (column:HUAPU C8 Extreme BDS 150 mm×30 mm×5 μm; mobile phase: [A, water (10 mMNH₄HCO₃)—B, ACN]; B %:35%-55%, 10 min). EXAMPLE 21,5-(6-chloroindolin-1-yl)sulfonyl-4-fluoro-isoquinolin-1-ol (5 mg, 12.2μmol, 6.61% yield) was obtained as a white solid. LC-MS: [M+1] 378.81.

¹H NMR (400 MHz, MeOD): δ 9.10 (s, 1H), 8.74 (d, J=7.6 Hz, 1H), 8.23 (d,J=7.6 Hz, 1H), 7.97 (d, J=6.8 Hz, 1H), 7.81 (t, J=7.6 Hz, 1H), 7.35 (d,J=8 Hz, 1H), 7.12˜7.15 (m, 1H), 4.16 (t, J=8.8 Hz, 2H), 3.23 (t, J=8.4Hz, 2H). One proton was exchanged.

Example 22

1-((1-Hydroxy-4-methylisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile

Compound 22-1: To a mixture of KNO₃ (21.3 g, 211.4 mmol) in H₂SO₄ (600mL) was added 4-bromoisoquinoline (40 g, 192.2 mmol) at 0° C. and thenthe mixture was stirred at 25° C. for 12 h under an N₂ atmosphere. Thereaction mixture was quenched by the addition of ice water (3000 mL) andthen NH₃.H₂O was added to the mixture to adjust to pH 8. The mixture wasfiltered and concentrated under reduced pressure to give a residue. Theresidue was washed by MeOH (500 mL) and then filtered. The solid wasdissolved with EtOAc (500 mL) and H₂O (1000 mL). The organic layer wasconcentrated under reduced pressure to give a residue. Compound 22-1,4-bromo-5-nitro-isoquinoline (75 g, 296.3 mmol, 77.0% yield), wasobtained as a yellow solid.

Compound 22-2: To a mixture of 4-bromo-5-nitro-isoquinoline (55 g, 217.3mmol) in dioxane (500 mL) and H₂O (50 mL) was added methylboronic acid(26.02 g, 434.6 mmol), Cs₂CO₃ (141.6 g, 434.6 mmol) and Pd(dppf)Cl₂(15.90 g, 21.73 mmol) in one portion. The mixture was heated to 110° C.and stirred for 12 h. The reaction mixture was filtered. The filtratewas diluted with H₂O (1.2 L) and extracted with EtOAc (800 mL×2). Thecombined organic layers were washed with brine (1.2 L), dried overNa₂SO₄, filtered and concentrated under reduced pressure to give aresidue. The residue was purified by column chromatography (SiO₂,petroleum ether:ethyl acetate=1:0 to 3:1). Compound 22-2,4-methyl-5-nitro-isoquinoline (36 g, 191.3 mmol, 88.0% yield), wasobtained as a yellow solid.

Compound 22-3: To a solution of 4-methyl-5-nitro-isoquinoline (33 g,175.3 mmol) in THF (500 mL) and H₂O (250 mL) was added Zn (91.7 g, 1.40mol) and NH₄Cl (75.0 g, 1.40 mol). The mixture was stirred at 25° C. for12 h. The reaction mixture was filtered and concentrated under reducedpressure to give a residue. The residue was purified by columnchromatography (SiO₂, petroleum ether:ethyl acetate=1:0 to 0:1).Compound 22-3, 4-methylisoquinolin-5-amine (20 g, 113.8 mmol, 64.8%yield), was obtained as a yellow solid.

Compound 22-4: Solution 1: To a mixture of 4-methylisoquinolin-5-amine(3 g, 18.9 mmol) in HCl (30 mL) was added NaNO₂ (1.31 g, 18.9 mmol) andH₂O (6 mL) in one portion at −5° C. under N₂. The mixture was stirred at−5° C. for 1 h. Solution 2: SO₂ was bubbled into a solution of CuCl(469.3 mg, 4.74 mmol) in HOAc (30 mL) and H₂O (6 mL) at 0° C. for 30minutes. The two solutions were mixed and stirred at 25° C. for 1.5 h.The reaction mixture was concentrated under reduced pressure to removesolvent. The residue was diluted with H₂O (300 mL) and extracted withDCM (200 mL×3). The combined organic layers were washed with brine (300mL), dried over Na₂SO₄, filtered and concentrated under reduced pressureto give a residue. Compound 22-4, 4-methylisoquinoline-5-sulfonylchloride (2 g, crude), was obtained as a yellow solid and was useddirectly without purification.

Compound 22-5: To a solution of 4-methylisoquinoline-5-sulfonyl chloride(2 g, 8.27 mmol) in acetone (30 mL) was added K₂CO₃ (3.43 g, 24.8 mmol)and indoline-6-carbonitrile INDOLINE 4 (1.19 g, 8.27 mmol). The mixturewas stirred at 25° C. for 12 h. The mixture was heated to 50° C. andstirred for 2 hours. The reaction mixture was diluted with H₂O (200 mL)and extracted with DCM (200 mL×2). The combined organic layers weredried over Na₂SO₄, filtered and concentrated under reduced pressure togive a residue. The residue was purified by column chromatography (SiO₂,petroleum ether:ethyl acetate=1:0 to 0:1). Compound 22-5,1-[(4-methyl-5-isoquinolyl)sulfonyl]indoline-6-carbonitrile (1.1 g, 2.52mmol, 30.4% yield), was obtained as a yellow solid.

Compound 22-6: To a solution of1-[(4-methyl-5-isoquinolyl)sulfonyl]indoline-6-carbonitrile (1.1 g, 3.15mmol) in DCM (30 mL) was added m-CPBA (1.44 g, 6.93 mmol) at 0° C. Themixture was warmed to 25° C. and stirred for 12 h. The reaction mixturewas purified directly by column chromatography (SiO₂, petroleumether:ethyl acetate=1:0, ethyl acetate: EtOH=1:1). Compound 22-6,1-(4-methyl-2-oxido-isoquinolin-2-ium-5-yl)sulfonylindoline-6-carbonitrile(1 g, 2.54 mmol, 80.7% yield), was obtained as a yellow solid.

EXAMPLE 22: A mixture of1-(4-methyl-2-oxido-isoquinolin-2-ium-5-yl)sulfonylindoline-6-carbonitrile(1 g, 2.74 mmol) in Ac₂O (20 mL) was heated to 120° C. and stirred for12 h. The reaction mixture was concentrated under reduced pressure toremove solvent. The residue was dissolved in DMF (20 mL) and filtered.The filtrate was purified by prep-HPLC (column: Waters Xbridge Prep OBDC18 150 mm×40 mm×10 rpm; mobile phase: [A, water (10 mM NH₄HCO₃)—B,ACN];B %: 20%-50%, 11 min) and concentrated. The product was added toEtOH (2 mL) and filtered to provide1-[(1-hydroxy-4-methyl-5-isoquinolyl)sulfonyl]indoline-6-carbonitrile(19 mg) as a light yellow solid. LC-MS: [M+1]365.08.

¹H NMR (400 MHz, DMSO-d₆): δ 11.71 (br, 1H), 8.59 (d, J=6.4 Hz, 1H),7.89 (d, J=6.4 Hz, 1H), 7.59˜7.51 (m, 3H), 7.25 (s, 2H), 4.22 (t, J=8.8Hz, 2H), 3.4 (t, J=8.4 Hz, 2H), 2.61 (s, 3H).

Example 23

1-((1-Hydroxy-4-methylisoquinolin-5-yl)sulfonyl)-4-methylindoline-6-carbonitrile

Compound 23-1: To a solution of 4-methylisoquinoline-5-sulfonyl chloride(100 mg, 413.7 μmol) in acetone (5 mL) was added K₂CO₃ (171.5 mg, 1.24mmol) and 4-methylindoline-6-carbonitrile INDOLINE 16 (52.36 mg, 331.0μmol). The mixture was stirred at 25° C. for 12 h. The mixture washeated to 50° C. and stirred for 6 hours. The reaction mixture wasdiluted with H₂O (100 mL) and extracted with DCM (100 mL×2). Thecombined organic layers were dried over Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by column chromatography (SiO₂, petroleum ether:ethylacetate=1:0 to 0:1). Compound 23-1,4-methyl-1-[(4-methyl-5-isoquinolyl)sulfonyl]indoline-6-carbonitrile (50mg, crude), was obtained as a yellow solid.

Compound 23-2: To a solution of4-methyl-1-[(4-methyl-5-isoquinolyl)sulfonyl]indoline-6-carbonitrile (50mg, 137 μmol) in DCM (10 mL) was added m-CPBA (62.9 mg, 302.7 μmol) at0° C. The mixture was heated to 25° C. and stirred for 2 h. The residuewas purified by column chromatography (SiO₂, petroleum ether:ethylacetate=1:0, ethyl acetate: MeOH=5:1). Compound 23-2,4-methyl-1-(4-methyl-2-oxido-isoquinolin-2-ium-5-yl)sulfonyl-indoline-6-carbonitrile(40 mg, crude), was obtained as a yellow solid.

EXAMPLE 23: A mixture of4-methyl-1-(4-methyl-2-oxido-isoquinolin-2-ium-5-yl)sulfonyl-indoline-6-carbonitrile(40 mg, 105.4 μmol) in Ac₂O (5 mL) was heated to 100° C. and stirred for12 h. The reaction mixture was concentrated under reduced pressure toremove solvent. The residue was purified by prep-HPLC (column: Luna C18100 mm×30 mm×5 m; mobile phase: [A, water (0.1% TFA)-B, ACN]; B %:30%-55%, 12 min). EXAMPLE 23, 1-[(1-hydroxy-4-methyl-5-isoquinolyl)sulfonyl]-4-methyl-indoline-6-carbonitrile (1.6 mg, 3.97 mol, 3.76%yield) was obtained as a brown solid. LC-MS: [M+1] 379.10.

¹H NMR (400 MHz, DMSO-d₆): δ 11.71 (br, 1H), 8.59 (d, J=8 Hz, 1H), 7.88(d, J=7.6 Hz, 1H), 7.53 (t, J=8 Hz, 1H), 7.39 (s, 1H), 7.26˜7.25 (m,1H), 7.09 (s, 1H), 4.22 (t, J=8 Hz, 2H) 2.61 (s, 6H) (2H may beoverlapped with the water peak).

Example 24

5-((6-Chloroindolin-1-yl)sulfonyl)-4-methylisoquinolin-1-ol

Compound 24-1: To a solution of 4-methylisoquinoline-5-sulfonyl chloride(500 mg, 2.07 mmol) in acetone (10 mL) was added K₂CO₃ (857.7 mg, 6.21mmol) and 6-chloroindoline INDOLINE 2 (317.8 mg, 2.07 mmol). The mixturewas stirred at 25° C. for 12 h. The reaction mixture was filtered andthe filtrate was diluted with H₂O (200 mL) and extracted with EtOAc (150mL×2). The combined organic layers were dried over Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by column chromatography (SiO₂, petroleum ether:ethylacetate=1:0 to 0:1). Compound 24-1,5-(6-chloroindolin-1-yl)sulfonyl-4-methyl-isoquinoline (400 mg, 947.4μmol, 45.80% yield), was obtained as a yellow solid.

Compound 24-2: To a solution of5-(6-chloroindolin-1-yl)sulfonyl-4-methyl-isoquinoline (400 mg, 1.11mmol) in DCM (10 mL) was added m-CPBA (579.3 mg, 2.79 mmol) at 0° C. Themixture was heated to 25° C. and stirred for 2.5 hours. The reactionmixture was purified directly by column chromatography (SiO₂, petroleumether:ethyl acetate=1:0, ethyl acetate:MeOH=5:1). Compound 24-2,5-(6-chloroindolin-1-yl)sulfonyl-4-methyl-2-oxido-isoquinolin-2-ium (300mg, crude), was obtained as a yellow solid.

EXAMPLE 24: A mixture of5-(6-chloroindolin-1-yl)sulfonyl-4-methyl-2-oxido-isoquinolin-2-ium (300mg, 800.3 μmol) in Ac₂O (10 mL) was heated to 100° C. and stirred for 12h. The reaction mixture was concentrated under reduced pressure toremove solvent. The residue was purified by prep-HPLC (column: WatersXbridge Prep OBD C18 150 mm×40 mm×10 μm; mobile phase: [A, water (10 mMNH₄HCO₃)—B, ACN]; B %: 30%-55%, 11 min). MeOH (2 mL) was added, themixture was filtered and concentrated under reduced pressure to give thecompound 24, 5-(6-chloroindolin-1-yl)sulfonyl-4-methyl-isoquinolin-1-ol(6 mg, 15.2 mol, 40.7% yield) as a yellow solid. LC-MS: [M+1] 374.05.

¹H NMR (400 MHz, DMSO-d₆): δ 11.70 (br, 1H), 8.59 (d, J=4.4 Hz, 1H),7.86 (d, J=7.2 Hz, 1H), 7.54 (t, J=7.6 Hz, 1H), 7.37 (d, J=8 Hz, 1H),7.25 (d, J=4.8 Hz 1H), 7.09 (d, J=8.8 Hz, 1H), 6.98 (s, 1H), 4.2 (t, J=8Hz, 1H), 3.26 (m, 2H, overlapped with solvent peak), 2.61 (s, 3H). Oneproton was exchanged.

Example 25

5-((6-Chloroindolin-1-yl)sulfonyl)phthalazin-1(2H)-one

Compound 25-2: A mixture of 4-nitroisoindoline-1,3-dione (25 g, 130.1mmol), NaBH₄ (11.13 g, 294.0 mmol) in MeOH (250 mL) was degassed andpurged with N₂ 3 times. The mixture was stirred at 25° C. for 2 h underan N₂ atmosphere. The solution was acidified with 20% HCl (100 mL) andthe reaction mixture was concentrated under reduced pressure to give aresidue. The dried residue was treated with acetone. The residue waspurified by column chromatography (SiO₂, petroleum ether:ethylacetate=10:1 to 0:1). Compound 25-2, 3-hydroxy-4-nitro-isoindolin-1-one(15 g, 59% yield) was obtained as a yellow solid.

Compound 25-3: A mixture of 3-hydroxy-4-nitro-isoindolin-1-one (10 g,51.5 mmol) in HCl (70 mL, 20% aq. solution) was degassed and purged withN₂ 3 times. The mixture was stirred at 90° C. for 12 h under an N₂atmosphere. The reaction mixture was concentrated under reduced pressureto give a residue. The residue was purified by column chromatography(SiO₂, petroleum ether:ethyl acetate=10:1 to 1:1). Compound 25-33-hydroxy-4-nitro-3H-isobenzofuran-1-one (10 g, crude) was obtained as ayellow solid and was used without further purification.

Compound 25-4: A mixture of 3-hydroxy-4-nitro-3H-isobenzofuran-1-one (10g, 51.2 mmol), N₂H₄.H₂O (9.05 g, 153.7 mmol) in EtOH (200 mL) wasdegassed and purged with N₂ 3 times. The mixture was stirred at 80° C.for 5 h under an N₂ atmosphere. The reaction mixture was concentratedunder reduced pressure to give a residue. The residue was purified bycolumn chromatography (SiO₂, petroleum ether:ethyl acetate toDCM:MeOH=10:1 to 10:1).

Compound 25-4, 5-nitro-2H-phthalazin-1-one (6 g, 31.3 mmol, 61.2% yield)was obtained as a yellow solid.

Compound 25-5: A mixture of 5-nitro-2H-phthalazin-1-one (6 g, 31.3mmol), Pd/C (3 g, 10 wt %) in MeOH (100 mL) was degassed and purged withH₂ 3 times, and then the mixture was stirred at 25° C. for 12 h under H₂atmosphere with a pressure of 15 psi. The reaction mixture was filteredand the filtrate was concentrated. The residue was purified by columnchromatography (SiO₂, petroleum ether:ethyl acetate to DCM:MeOH=10:1 to10:1). Compound 25-5, 5-amino-2H-phthalazin-1-one (3 g, 18.6 mmol, 59.3%yield) was obtained as a yellow solid.

Compound 25-6: To a solution of 5-amino-2H-phthalazin-1-one (3 g, 18.61mmol) in HCl (20 mL) was added NaNO₂ (1.28 g, 18.61 mmol) and H₂O (5 mL)at −5° C. The mixture was stirred at −5° C. for 1 h. Then it wastransferred in one portion to a solution of AcOH (15 mL) with CuCl(460.7 mg, 4.65 mmol) and H₂O (5 mL) that was saturated by bubbling SO₂.The mixture was stirred at 25° C. for 2 h under an N₂ atmosphere. Thereaction mixture was concentrated under reduced pressure to give aresidue. The crude product of Compound 25-6,1-oxo-2H-phthalazine-5-sulfonyl chloride (1.5 g, 6.13 mmol, 32.9%yield), was obtained as a yellow solid and was used in the next stepwithout further purification.

EXAMPLE 25: To a solution of 1-oxo-2H-phthalazine-5-sulfonyl chloride (1g, 4.09 mmol) in DCM (15 mL) was added TEA (853.3 L L, 6.13 mmol) and6-chloroindoline INDOLINE 2 (502.2 mg, 3.27 mmol). The mixture wasstirred at 25° C. for 12 h under an N₂ atmosphere. The reaction mixturewas diluted with H₂O (50 mL) and extracted with DCM (150 mL×2). Thecombined organic layers were washed with saturated brine (10 mL), driedover anhydrous Na₂SO₄, filtered and concentrated under reduced pressureto give a residue. The residue was purified by column chromatography(SiO₂, petroleum ether:ethyl acetate=10:1 to 0:1). Compound 25,5-(6-chloroindolin-1-yl)sulfonyl-2H-phthalazin-1-one (0.15 g, 373.3μmol, 9.13% yield) was obtained as a yellow solid. LC-MS: [M+1] 361.03.

¹H NMR (400 MHz, DMSO-d₆): δ 13.08 (s, 1H), 8.78 (s, 1H), 8.56 (d, J=8Hz, 1H), 8.46 (d, J=8 Hz, 1H), 8.02 (t, J=8 Hz, 1H), 7.39 (s, 1H), 7.22(d, J=8.4 Hz, 1H), 7.09 (d, J=8.4 Hz, 1H), 4.06 (t, J=8 Hz, 2H), 2.90(t, J=8.4 Hz, 2H).

Example 26

1-((1-Oxo-1,2-dihydrophthalazin-5-yl)sulfonyl)indoline-6-carbonitrile

EXAMPLE 26: To a solution of 1-oxo-2H-phthalazine-5-sulfonyl chloride(0.5 g, 2.04 mmol) in DCM (15 mL) was added TEA (426 μL, 3.07 mmol) andindoline-6-carbonitrile INDOLINE 4 (235 mg, 1.63 mmol). The mixture wasstirred at 25° C. for 12 h under an N₂ atmosphere. The reaction mixturewas diluted with H₂O (20 mL) and extracted with EtOAc (80 mL×2). Thecombined organic layers were washed with saturated brine (20 mL), driedover anhydrous Na₂SO₄, filtered and concentrated under reduced pressureto give a residue. The residue was purified by column chromatography(SiO₂, petroleum ether:ethyl acetate=10:1 to 0:1). Compound 26,1-[(1-oxo-2H-phthalazin-5-yl)sulfonyl]indoline-6-carbonitrile (30 mg,81.6 mol, 3.99% yield) was obtained as a yellow solid. LC-MS: [M+1]352.06.

¹H NMR (400 MHz, DMSO-d₆): δ 13.09 (s, 1H), 8.81 (s, 1H), 8.56 (d, J=8Hz, 2H), 8.01 (t, J=8 Hz, 1H), 7.72 (s, 1H), 7.52 (d, J=9.2 Hz, 1H),7.41 (d, J=7.6 Hz, 1H), 4.08 (t, J=8.4 Hz, 2H), 3.03 (t, J=8.4 Hz, 2H).

Example 27

1-((4-chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile

Compound 27-2. To a mixture of 4-chloroisoquinoline (5 g, 30.56 mmol, 1eq) in H₂SO₄ (20 mL) KNO₃ (3.40 g, 33.62 mmol, 1.1 eq) was added at 0°C. under N₂. The mixture was stirred at 15° C. for 12 hours. Thereaction mixture was added to ice-cold water (100 ml), and thensaturated NaOH solution was added until the pH was adjusted to pH=10.The mixture was extracted with EtOAc (200 mL×2). The combined organiclayers were washed with brine (200 mL), dried over Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue. The residue waswashed with the mixture solvent of petroleum ether:ethyl acetate=1:1 (50mL). The mixture was filtered and concentrated under reduced pressure togive a residue. Compound 27-2, 4-chloro-5-nitro-isoquinoline (4 g, 19.18mmol, 62.74% yield), was obtained as a yellow solid.

Compound 27-3: To a solution of 4-chloro-5-nitro-isoquinoline (4 g,19.18 mmol, 1 eq) and NH₄Cl (8.21 g, 153.40 mmol, 8 eq) in THF (50 mL)and H₂O (25 mL) Zn (10.03 g, 153.40 mmol, 8 eq) was added at 0° C. Theresulting mixture was stirred at 15° C. for 3 hr. The reaction mixturewas filtered, and the filtrate was added to H₂O (200 mL) and extractedwith EtOAc (150 mL×2). The combined organic layers were washed withbrine (200 mL), dried over Na₂SO₄, filtered and concentrated underreduced pressure to give a residue. The residue was purified by columnchromatography (SiO₂, petroleum ether:ethyl acetate=1:0 to 0:1).Compound 27-3, 4-chloroisoquinolin-5-amine (1.5 g, 8.40 mmol, 43.79%yield), was obtained as a light yellow solid.

Preparation of Solution 1: To a mixture of 4-chloroisoquinolin-5-amine(1.5 g, 8.40 mmol, 1 eq) in conc. HCl (15 mL) was added NaNO2 (579.45mg, 8.40 mmol, 1 eq) in one portion at −5° C. under N₂. The mixture wasstirred at −5° C. for 1 hr.

Preparation of Solution 2: SO₂ was bubbled into a solution of CuCl(207.85 mg, 2.10 mmol, 0.25 eq) in HOAc (15 mL) and H₂O (3 mL) at 0° C.for 30 minutes.

Compound 27-4: To solution 1 solution 2 was added slowly. The mixturewas stirred at 15° C. for 2 hr. The reaction mixture was concentratedunder reduced pressure to remove solvent.

The residue was diluted with H₂O (500 mL) and extracted with DCM (300mL×3). The combined organic layers were washed with brine (500 mL),dried over Na₂SO₄, filtered and concentrated under reduced pressure togive a residue that was used with no further purification. Compound27-4, 4-chloroisoquinoline-5-sulfonyl chloride (1.5 g, crude), wasobtained as a yellow solid.

Compound 27-5: To a mixture of 4-chloroisoquinoline-5-sulfonyl chloride(1.5 g, 5.72 mmol, 1 eq) and indoline-6-carbonitrile INDOLINE 4 (825.06mg, 5.72 mmol, 1 eq) in DMF (20 mL) K₂CO₃ (1.58 g, 11.45 mmol, 2 eq) wasadded in one portion at 15° C. under N₂. The mixture was stirred at 15°C. for 12 hours. The reaction mixture was diluted with H₂O (200 mL) andextracted with EtOAc (150 mL×3). The combined organic layers were driedover Na₂SO₄, filtered and concentrated under reduced pressure to give aresidue. The residue was purified by column chromatography (SiO₂,petroleum ether:ethyl acetate=1:0 to 0:1). Compound 27-5,1-[(4-chloro-5-isoquinolyl)sulfonyl]indoline-6-carbonitrile (600 mg,crude), was obtained as a yellow solid.

Compound 27-6: To a solution of1-[(4-chloro-5-isoquinolyl)sulfonyl]indoline-6-carbonitrile (600 mg,1.62 mmol, 1 eq) in DCM (20 mL) was added m-CPBA (769.93 mg, 3.57 mmol,80% purity, 2.2 eq).The mixture was stirred at 15° C. for 12 hr. Thereaction mixture was purified directly by column chromatography (SiO₂,petroleum ether:ethyl acetate=1:0 to ethyl acetate: EtOH=3:1). The crudeproduct of Compound 27-6 was obtained triturated with MeOH (50 mL),filtered and was used in the next step without further purification.

EXAMPLE 27: A mixture of1-(4-chloro-2-oxido-isoquinolin-2-ium-5-yl)sulfonylindoline-6-carbonitrile(150 mg, 388.78 μmol, 1 eq) in Ac₂O (15 mL) was heated to 100° C. andstirred for 12 hours. The reaction mixture was concentrated underreduced pressure to remove solvent. The residue was purified byprep-HPLC (column: Xtimate C18 150×25 mm×5 μm; mobile phase: [water(10mM NH₄HCO₃)-ACN];B %: 40%-70%,10 min). Then the residue was re-purifiedby prep-HPLC (column: Xtimate C18 150×25 mm×5 μm;mobile phase: [water(10mM NH₄HCO₃)-ACN];B %: 30%-55%, 8 min). EXAMPLE 27,1-[(4-chloro-1-oxo-2H-isoquinolin-5-yl)sulfonyl]indoline-6-carbonitrile(4 mg, 9.63 μmol, 2.48% yield, 92.93% purity), was obtained as a yellowsolid. LCMS (ESI+) m/z: 383.0.

¹H NMR (400 MHz, DMSO-d₆): 8.55 (d, J=8 Hz, 1H), 7.98 (d, J=7.6 Hz, 1H),7.63-7.46 (m, 4H), 7.21 (s, 1H), 4.15 (t, J=8.6 Hz, 2H), 3.34 (t, J=8.8Hz, 2H).

Example 28

1-((1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)-2,3-dihydro-1H-pyrrolo[3,2-c]pyridine-6-carbonitrile

Compound 28-2: To a solution of 6-chloro-2,3-dihydro-1H-pyrrolo[3,2-c]pyridine (1.6 g, 10.35 mmol, 1 eq) in DCM (20mL), Boc₂O (6.78 g, 31.05 mmol, 7.13 mL, 3 eq), TEA (2.09 g, 20.70 mmol,2.88 mL, 2 eq) and DMAP (1.26 g, 10.35 mmol, 1 eq) were added at 20° C.The mixture was heated to 50° C. and stirred for 12 hours. The reactionmixture was concentrated under reduced pressure to remove solvent. Theresidue was purified by column chromatography (SiO₂, petroleumether:ethyl acetate=1:0 to 0:1). Compound 28-2, tert-butyl 6-chloro-2,3-dihydropyrrolo[3,2-c]pyridine-1-carboxylate (1.5 g, 5.89 mmol, 56.90%yield), was obtained as a colorless oil.

Compound 28-3: To a solution of tert-butyl6-chloro-2,3-dihydropyrrolo[3,2-c]pyridine-1-carboxylate (1.5 g, 5.89mmol, 1 eq) in MeOH (50 mL) and DIPEA (5 mL) Pd(dppf)Cl₂.CH₂Cl₂ (2.40 g,2.94 mmol, 0.5 eq) was added at 15° C. The suspension was degassed undervacuum and purged with CO several times. The mixture was stirred underCO (50 psi) at 80° C. for 12 hours. The reaction mixture wasconcentrated under reduced pressure to remove solvent. The residue waspurified by column chromatography (SiO₂, petroleum ether:ethylacetate=1:0 to 0:1). Compound 28-3, Ol-tert-butyl 06-methyl2,3-dihydropyrrolo[3,2-c]pyridine-1,6-dicarboxylate (1.5 g, 4.31 mmol,73.22% yield, 80% purity), was obtained as a red solid.

Compound 28-4: To a solution of Ol-tert-butyl 06-methyl2,3-dihydropyrrolo[3,2-c]pyridine-1,6-dicarboxylate (1.57 g, 5.64 mmol,1 eq) in MeOH (20 mL) and H₂O (4 mL) NaOH (676.96 mg, 16.92 mmol, 3 eq)was added. The mixture was stirred at 15° C. for 30 min.The reactionmixture was concentrated under reduced pressure to remove MeOH. Then tothe remaining solution was added 12M HCl solution until pH=3˜4, and themixture was extracted with EtOAc (100 mL×2). The combined organic layerswere dried over Na₂SO₄, filtered and concentrated under reduced pressureto give a residue that was used with no further purification. Compound28-4,1-tert-butoxycarbonyl-2,3-dihydropyrrolo[3,2-c]pyridine-6-carboxylicacid (700 mg, crude), was obtained as a yellow solid.

Compound 28-5: To a solution of1-tert-butoxycarbonyl-2,3-dihydropyrrolo[3,2-c]pyridine-6-carboxylicacid (650 mg, 2.46 mmol, 1 eq) in DMF (10 mL) HATU (1.40 g, 3.69 mmol,1.5 eq), NH₄Cl (526.24 mg, 9.84 mmol, 4 eq) and DIPEA (1.59 g, 12.30mmol, 2.14 mL, 5 eq) were added. The mixture was stirred at 15° C. for12 hr. The reaction mixture was added to H₂O (150 mL) and extracted withEtOAc (80 mL×2). The combined organic layers were washed with brine (150mL), dried over Na₂SO₄, filtered and concentrated under reduced pressureto give a residue. The residue was purified by column chromatography(SiO₂, petroleum ether:ethyl acetate=1:0 to 0:1). Compound 28-5,tert-Butyl 6-carbamoyl-2,3-dihydropyrrolo[3,2-c]pyridine-1-carboxylate(300 mg, 46% yield), was obtained as a white solid.

Compound 28-6: To a solution of tert-butyl6-carbamoyl-2,3-dihydropyrrolo[3,2-c]pyridine-1-carboxylate (250 mg,949.52 μmol, 1 eq) in THF (10 mL), pyridine (195.28 mg, 2.47 mmol,199.26 μL, 2.6 eq) and TFAA (518.51 mg, 2.47 mmol, 343.38 μL, 2.6 eq)were added. The mixture was stirred at 15° C. for 2 hr. The reactionmixture was added to H₂O (100 mL) and extracted with EtOAc (50 mL×2).The combined organic layers were washed with brine (100 mL), dried overNa₂SO₄, filtered and concentrated under reduced pressure to give aresidue. The residue was purified by column chromatography (SiO₂,petroleum ether:ethyl acetate=1:0 to 1:1). Compound 28-6, tert-butyl6-cyano-2,3-dihydropyrrolo[3,2-c]pyridine-1-carboxylate (250 mg, crude),was obtained as a light yellow solid.

Compound 28-7: A solution of tert-butyl6-cyano-2,3-dihydropyrrolo[3,2-c]pyridine-1-carboxylate (250 mg, 1.02mmol, 1 eq) in HCl:EtOAc (10 mL) was stirred at 15° C. for 12 hr. Thereaction mixture was concentrated under reduced pressure to removesolvent. The residue was used with no further purification. Compound28-7, 2,3-dihydro-1H-pyrrolo[3,2-c]pyridine-6-carbonitrile (160 mg,crude, HCl), was obtained as an off-white solid.

Compound 1-4: A mixture of 1-hydroxyisoquinoline-5-sulfonic acid 1-3(1.00 g, 4.44 mmol, 1 eq) in SOCl₂ (10 mL) and DMF (0.1 mL) was heatedto 40° C. and stirred for 1 h. The reaction mixture was concentratedunder reduced pressure to remove the solvent. The residue was dilutedwith H₂O (100 mL) and extracted with DCM (70 mL×2). The combined organiclayers were washed with brine (100 mL), dried over Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by column chromatography (SiO₂, petroleum ether:ethylacetate=1:0 to 0:1). Compound 1-4,1-oxo-2H-isoquinoline-5-sulfonylchloride (300 mg, crude) was obtained as a yellow solid which was useddirectly in the next step.

EXAMPLE 28: To a solution of2,3-dihydro-1H-pyrrolo[3,2-c]pyridine-6-carbonitrile 28-7 (37.53 mg,258.55 μmol, 0.7 eq) in DMF (7 mL) NaH (59.10 mg, 1.48 mmol, 78.02 μL,60% purity, 4 eq) was added at 15° C. The mixture was stirred at thistemperature for 30 min, and then 1-oxo-2H-isoquinoline-5-sulfonylchloride 1-4 (90 mg, 369.36 μmol, 1 eq) was added at 15° C. Theresulting mixture was stirred at 15° C. for 1.5 hr. The reaction mixturewas quenched by addition of H₂O (20 mL), and then diluted with H₂O (40mL) and extracted with EtOAc (20 mL×2). The combined organic layers werewashed with brine (60 mL), dried over Na₂SO₄, filtered and concentratedunder reduced pressure to give a residue. The residue was purified byprep-HPLC (column: Xtimate C18 150×25 mm×5 m; mobile phase: [water(10 mMNH₄HCO₃)-ACN];B %: 20%-50%, 8 min). EXAMPLE 28,1-[(1-oxo-2H-isoquinolin-5-yl)sulfonyl]-2,3-dihydropyrrolo[3,2-c]pyridine-6-carbonitrile(3 mg, purity 98.76%), was obtained as a white solid. LCMS (ESI+)/mz:353.0.

¹H NMR (400 MHz, DMSO-d₆): δ 11.74 (br, 1H), 8.56 (d, J=8 Hz, 1H),8.51˜8.46 (m, 2H), 7.75 (s, 1H), 7.67 (t, J=8 Hz, 1H), 7.45˜7.37 (m,1H), 7.11 (d, J=7.6 Hz, 1H), 4.10 (t, J=8.8 Hz, 2H), 3.22 (t, J=8.6 Hz,2H).

Example 29

4-methoxy-1-((1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile

Compound 29-2: To a solution of 4-methoxy-1H-indole-6-carbonitrile (230mg, 1.34 mmol, 1 eq) in DCM (8 mL) was added triethylsilane (776.63 mg,6.68 mmol, 1.07 mL, 5 eq). The reaction was then cooled to 0° C. and TFA(4 mL) was added dropwise at 0° C. The mixture was heated to 50° C. andstirred for 12 hours. The reaction mixture was concentrated underreduced pressure to remove solvent. The residue was purified by columnchromatography (SiO₂, petroleum ether:ethyl acetate=1:0 to 0:1) Compound29-2, 4-methoxyindoline-6-carbonitrile (100 mg, 40% yield), was obtainedas a light yellow solid.

EXAMPLE 29: To a solution of 1-oxo-2H-isoquinoline-5-sulfonyl chloride1-4 (150 mg, 615.60 μmol, 1 eq) and 4-methoxyindoline-6-carbonitrile29-2 (32.17 mg, 184.68 μmol, 0.3 eq) in DCM (10 mL) was added TEA(207.64 mg, 1.23 mmol, 285.61 μL, 60% purity, 2 eq). The mixture wasstirred at 15° C. for 12 hr. The reaction mixture was added to H₂O (50mL) and extracted with EtOAc (30 mL×2). The combined organic layers wereconcentrated under reduced pressure to give a residue. The residue waspurified by prep-HPLC (column: Nano-micro Kromasil C18 100×40 mm 10 μm;mobile phase: [water(0.1% TFA)-ACN];B %: 30%-56%,10 min). The residuewas re-purified by prep-HPLC (column: Xtimate C18 150×25 mm×5 μm; mobilephase: [water(10 mM NH₄HCO₃)-ACN];B %: 25%-55%, 8 min). EXAMPLE 29,4-methoxy-1-[(1-oxo-2H-isoquinolin-5-yl)sulfonyl]indoline-6-carbonitrile(2 mg, 96.64% purity), was obtained as a white solid. LCMS (ESI+) m/z:382.0.

¹H NMR (400 MHz, DMSO-d₆): δ 11.68 (br, 1H), 8.52 (d, J=8 Hz, 1H), 8.34(d, J=7.6 Hz, 1H), 7.64 (t, J=8 Hz, 1H), 7.38 (d, J=7.6 Hz, 1H), 7.26(s, 1H), 7.21 (s, 1H), 7.13 (d, J=7.6 Hz, 1H), 4.07 (t, J=8.6 Hz, 2H),3.82 (s, 3H), 2.99 (t, J=8.6 Hz, 2H).

Examples 30 & 31

N-(6-cyano-1-((1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indolin-4-yl)acetamide

4-amino-1-((1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile

Compound 30-2: To a solution of 4-nitro-1H-indole-6-carbonitrile (1.00g, 5.34 mmol, 1 eq) in EtOH (20 mL) Pd/C (1.00 g, 106.86 μmol, 5% wt %)under N₂. The suspension was degassed under vacuum and purged with H₂several times. The mixture was stirred under H₂ (15 psi) at 15° C. for1.5 hours. The reaction mixture was filtered and the filtrateconcentrated under reduced pressure to give a residue that was used withno further purification. Compound 30-2, 4-amino-1H-indole-6-carbonitrile(700 mg, 4.45 mmol, 83.35% yield), was obtained as a yellow solid.

Compound 30-3: To a solution of 4-amino-1H-indole-6-carbonitrile (700mg, 4.45 mmol, 1 eq) in DCM (15 mL) was added TEA (1.35 g, 13.36 mmol,1.86 mL, 3 eq) and acetyl chloride (419.54 mg, 5.34 mmol, 381.40 μL, 1.2eq). The mixture was stirred at 15° C. for 2 hr. The reaction mixturewas concentrated under reduced pressure to remove solvent. The residuewas purified by column chromatography (SiO₂, petroleum ether:ethylacetate=1:0 to 0:1) by 2 times. Compound 30-3,N-(6-cyano-1H-indol-4-yl)acetamide (600 mg, 3.01 mmol, 67.63% yield),was obtained as a light yellow solid.

Compound 30-4: To a solution of N-(6-cyano-1H-indol-4-yl)acetamide (600mg, 3.01 mmol, 1 eq) in DCM (20 mL) TESH (3.50 g, 30.12 mmol, 4.81 mL,10 eq) was added. The reaction was then cooled to 0° C. and TFA (10 mL)was added dropwise at 0° C. The resulting mixture was stirred at 15° C.for 12 hr. The mixture was heated to 50° C. and stirred for 5 hours. Thereaction mixture was concentrated under reduced pressure to removesolvent. The residue was purified by column chromatography (SiO₂,petroleum ether:ethyl acetate=1:0 to 0:1). Compound 30-4,N-(6-cyanoindolin-4-yl)acetamide (400 mg, 1.99 mmol, 66.00% yield), wasobtained as a light yellow solid.

EXAMPLE 30: To a solution of 1-oxo-2H-isoquinoline-5-sulfonyl chloride1-4 (200 mg, 820.79 μmol, 1 eq) in DCM (10 mL), TEA (332.22 mg, 3.28mmol, 456.98 μL, 4 eq) and N-(6-cyanoindolin-4-yl)acetamide (82.58 mg,410.40 μmol, 0.5 eq) were added. The mixture was stirred at 15° C. for12 hr. The reaction mixture was added to H₂O (150 mL) and extracted withEtOAc (80 mL×2). The combined organic layers were concentrated underreduced pressure to give a residue. The residue was purified by columnchromatography (SiO₂, petroleum ether:ethyl acetate=1:0, DCM: MeOH=2:1).The residue was re-purified by prep-HPLC (column: Xtimate C18 150×25mm×5 m; mobile phase: [water(10 mM NH₄HCO₃)-ACN];B %: 20%-50%,8 min).EXAMPLE 30,N-[6-cyano-1-[(1-oxo-2H-isoquinolin-5-yl)sulfonyl]indolin-4-yl]acetamide(20 mg, 96.9% purity), was obtained as a white solid. LCMS (ESI+) m/z:409.1.

¹H NMR (400 MHz, DMSO-d₆): δ 11.67 (br, 1H), 9.58 (s, 1H), 8.49 (d, J=8Hz, 1H), 8.33 (d, J=8 Hz, 1H), 7.83 (s, 1H), 7.61 (t, J=8 Hz, 1H),7.36˜7.28 (m, 2H), 7.07 (d, J=7.2 Hz, 1H), 4.04 (t, J=8.4 Hz, 2H), 2.97(t, J=8.4 Hz, 2H), 2.03 (s, 1H).

EXAMPLE 31: A mixture ofN-[6-cyano-1-[(1-oxo-2H-isoquinolin-5-yl)sulfonyl]indolin-4-yl]acetamide(40 mg, 97.94 μmol, 1 eq) in conc. HCl (6 mL) and EtOH (6 mL) was heatedto 50° C. and stirred for 4.5 hours. The reaction mixture was added toaq. Na₂CO₃ until pH>7, and then diluted with H₂O (20 mL) and extractedwith DCM (50 mL×2).

The combined organic layers were concentrated under reduced pressure togive a residue. The residue was purified by prep-HPLC (column: WelchUltimate AQ-C18 150×30 mm×5 μm; mobile phase: [water(0.1% TFA)-ACN];B %:25%-55%,12 min). EXAMPLE 31,4-amino-1-[(1-oxo-2H-isoquinolin-5-yl)sulfonyl]indoline-6-carbonitrile(14 mg, 98.04% purity, TFA salt), was obtained as a white solid. LCMS(ESI+) m/z: 367.1.

¹H NMR (400 MHz, DMSO-d₆): δ 11.69 (d, J=5.6 Hz, 1H), 8.51 (d, J=7.6 Hz,1H), 8.27 (d, J=7.6 Hz, 1H), 7.63 (t, J=7.8 Hz, 1H), 7.37 (t, J=6.8 Hz,1H), 7.13 (d, J=7.6 Hz, 1H), 6.78 (s, 1H), 6.61 (s, 1H), 5.65 (br, 2H),4.05 (t, J=8.4 Hz, 2H), 2.77 (t, J=8.4 Hz, 2H).

Examples 32, 33 & 34

N-(6-cyano-1-((4-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indolin-4-yl)acetamide

4-amino-1-((4-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile

4-(dimethylamino)-1-((4-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile

Compound 22-1: Three reaction mixtures were prepared in parallel asfollows: to a solution of 4-bromoisoquinoline 21-1 (100 g, 480.64 mmol,1 eq) in H₂SO₄ (250 mL) KNO₃ (53.45 g, 528.71 mmol, 1.1 eq) was added at0° C. The mixture was stirred at 25° C. for 12 hr under N₂ atmosphere.The reaction mixture was quenched by addition to ice water (1 L) at 0°C., and then aq. NaOH solution (800 mL) was added to the mixture toadjust pH to −8. The mixture was filtered and the filter cake wasconcentrated under reduced pressure to give a residue. The residue wastriturated with petroleum ether: EtOAc=1:1.5 (2.5 L) at 25° C. for 12hr. Filtered and the solid was triturated with H₂O (1. 5 L) at 25° C.for 5 hr. Compound 32-2, 4-bromo-5-nitro-isoquinoline 22-1 (200 g,790.35 mmol, 54.81% yield), was obtained as a yellow solid. LCMS (ESI+)m/z: 252.9.

Compound 22-2: Two reaction mixtures were prepared in parallel asfollows: a mixture of 4-bromo-5-nitro-isoquinoline 22-1 (50 g, 197.59mmol, 1 eq), Cs₂CO₃ (128.76 g, 395.18 mmol, 2 eq), Pd(dppf)Cl₂ (14.46 g,19.76 mmol, 0.1 eq) and MeB(OH)₂ (23.66 g, 395.18 mmol, 2 eq) in dioxane(400 mL) and H₂O (40 mL) was degassed and purged with N₂ 3 times, andthen the mixture was stirred at 110° C. for 12 hr under N₂ atmosphere.Two batches with the same scale (50 g) were run in parallel and runwork-up/purification together. The reaction mixture was concentratedunder reduced pressure to remove dioxane. The residue was diluted withH₂O (1.5 L) and extracted with EtOAc (2 L×3). The combined organiclayers were dried over Na₂SO₄, filtered and concentrated under reducedpressure to give a residue. The residue was purified by columnchromatography (SiO₂, petroleum ether:ethyl acetate=10:1 to 1:1).Compound 22-2, 4-methyl-5-nitro-isoquinoline (40 g, 212.56 mmol, 53.79%yield), was obtained as a yellow solid.

Compound 32-4: To a solution of 4-methyl-5-nitro-isoquinoline 22-2 (60g, 318.84 mmol, 1 eq) in DCM (800 mL) was added m-CPBA (151.31 g, 701.44mmol, 80% purity, 2.2 eq) at 0° C. The mixture was stirred at 25° C. for12 hr. The reaction mixture was quenched by addition aq. Na₂SO₃ (900mL). The reaction mixture was diluted with H₂O (1 L) and extracted withDCM (2000 mL×4). The combined organic layers were dried over anhydrousNa₂SO₄, filtered and concentrated under reduced pressure to give aresidue. The residue was triturated with EtOAc (2000 mL) at 25° C. for 2hr and the solid was dried under reduced pressure. Compound 32-4,4-methyl-nitro-2-xido-isquinolin-2-ium (40 g, 195.90 mmol, 61.44%yield), was obtained as a yellow solid.

Compound 32-5: To a solution of4-methyl-5-nitro-2-oxido-isoquinolin-2-ium (45 g, 220.39 mmol, 1 eq) inDCM (500 mL), POCl₃ (40.55 g, 264.47 mmol, 24.58 mL, 1.2 eq) and DMF(8.05 g, 110.20 mmol, 8.48 mL, 0.5 eq) were added at 0° C. The mixturewas stirred at 20° C. for 12 hr under N₂ atmosphere. The reactionmixture was concentrated under reduced pressure to remove DCM. Theresidue was quenched with H₂O (300 mL) and then it was filtered. Thefilter cake was dried under reduced pressure to give a residue that wasused with no further purification. The crude product Compound 32-5,1-chloro-4-methyl-5-nitro-isoquinoline (27 g, crude), was obtained as ayellow solid. LCMS (ESI+) m/z: 222.9.

Compound 32-6: A mixture of 1-chloro-4-methyl-5-nitro-isoquinoline (27g, 121.28 mmol, 1 eq), NaOMe (13.10 g, 242.56 mmol, 2 eq) in MeOH (500mL) was degassed and purged with N₂ 3 times, and then the mixture wasstirred at 80° C. for 5 hr under a N₂ atmosphere. The reaction mixturewas concentrated under reduced pressure to remove MeOH. The residue wasdiluted with H₂O (500 mL) and extracted with EtOAc (600 mL×4). Thecombined organic layers were dried over Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by column chromatography (SiO₂, petroleum ether:ethylacetate=1:0 to 5:1). Compound 32-6,1-methoxy-4-methyl-5-nitro-isoquinoline (15 g, 68.74 mmol, 56.68%yield), was obtained as a yellow solid.

Compound 32-7: To a solution of 1-methoxy-4-methyl-5-nitro-isoquinoline(15 g, 68.74 mmol, 1 eq) in MeOH (210 mL) and H₂O (70 mL), Zn (22.48 g,343.71 mmol, 5 eq) and NH₄Cl (18.39 g, 343.71 mmol, 5 eq) were added at0° C. The mixture was stirred at 25° C. for 12 hr. The mixture wasfiltered and the filtrate was concentrated under reduced pressure togive a residue. The residue was purified by column chromatography (SiO₂,petroleum ether:ethyl acetate=10:1 to 1:1). Compound 32-7,1-methoxy-4-methyl-isoquinolin-5-amine (10 g, 53.13 mmol, 77.29% yield),was obtained as a brown solid. LCMS (ESI+) m/z: 189.0.

Compound 32-8: To a solution of 1-methoxy-4-methyl-isoquinolin-5-amine(2 g, 10.63 mmol, 1 eq) in HCl (80 mL), NaNO₂ (903.12 mg, 10.63 mmol, 1eq) in H₂O (5 mL) was added at −5° C. The mixture was stirred at −5° C.for 1 hr and transferred to a solution of AcOH (50 mL) with CuCl (262.98mg, 2.66 mmol, 63.52 μL, 0.25 eq) in H₂O (5 mL) which was saturated bySO₂ in one portion. The mixture was stirred at 20° C. for 2 hr. Thereaction mixture was quenched by addition to ice water (100 mL) at 0°C., and then the mixture was extracted with DCM (200 mL×3). The combinedorganic layers were dried over Na₂SO₄, filtered and concentrated underreduced pressure to give a residue that was used with no furtherpurification. Compound 32-8, 1-methoxy-4-methyl-isoquinoline-5-sulfonylchloride (1.5 g, crude), was obtained as a yellow solid. LCMS (ESI+)m/z: 271.9.

Compound 32-9: To a mixture of N-(6-cyanoindolin-4-yl)acetamide 30-4(888.67 mg, 4.42 mmol, 1.2 eq) and pyridine (1.05 g, 13.25 mmol, 1.07mL, 3.6 eq) was added dropwise a solution of1-methoxy-4-methylisoquinoline-5-sulfonyl chloride 32-8 (780 mg, 2.87mmol, 0.78 eq) in DCM (10 mL) −5° C. The mixture was degassed and purgedwith N₂ 3 times, and then the mixture was stirred at 15° C. for 12 hrunder N₂ atmosphere. The reaction mixture was filtered and concentratedunder reduced pressure to give a residue. Crude Compound 32-9,N-(6-cyano-1-((1-methoxy-4-methylisoquinolin-5-yl)sulfonyl)indolin-4-yl)acetamide(1.2 g, crude), was obtained as a brown solid and used into next stepwithout further purification.

EXAMPLE 32: A mixture ofN-(6-cyano-1-((1-methoxy-4-methylisoquinolin-5-yl)sulfonyl)indolin-4-yl)acetamide(600 mg, 1.37 mmol, 1 eq) and TMSI (1.10 g, 5.50 mmol, 748.43 μL, 4 eq)in ACN (10 mL) was stirred at 15° C. for 1 hr under N₂ atmosphere. Thereaction mixture was quenched by addition H₂O (30 mL) at 0° C., and thenextracted with EtOAc (30 mL×2). The combined organic layers were washedwith brine (60 mL×1), dried over Na₂SO₄, filtered and concentrated underreduced pressure to give a residue. The residue was purified by prep-TLC(SiO₂, DCM: MeOH=10:1), then it was re-purified by prep-HPLC (column:Waters Xbridge BEH C18 100×25 mm×5 m; mobile phase: [water (10 mMNH₄HCO₃)-ACN]; B %: 15%-50%, 8 min). EXAMPLE 32,N-(6-cyano-1-((4-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indolin-4-yl)acetamide(4 mg, 9.46 μmol, 99.9% purity), was obtained as a white solid. LCMS(ESI+) m/z: 423.1.

¹H NMR (400 MHz, DMSO-d₆): δ 11.69 (br, 1H), 9.74 (s, 1H), 8.59 (d, J=4Hz, 1H), 7.98˜7.85 (m, 2H), 7.52 (t, J=7.6 Hz, 1H), 7.24 (s, 1H), 6.99(s, 1H), 4.22 (t, J=8.4 Hz, 2H), ˜3.31 (2H, mixed with solvent peak),2.59 (s, 3H), 2.14 (s, 3H).

EXAMPLE 33: A solution ofN-(6-cyano-1-((4-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indolin-4-yl)acetamide(30 mg, 71.01 μmol, 1 eq) in conc. HCl (5 mL) and EtOH (5 mL) wasstirred at 50° C. for 5 hr. The reaction mixture was added saturatedNa₂CO₃ solution until pH=8 at 0° C., and then extracted with EtOAc (60mL×5). The combined organic layers were washed with brine (200 mL×1),dried over Na₂SO₄, filtered and concentrated under reduced pressure togive a residue. The residue was purified by prep-HPLC (column: WatersXbridge BEH C18 100×25 mm×5 m; mobile phase: [water (10 mMNH₄HCO₃)-ACN]; B %: 20%-50%, 8 min). EXAMPLE 33,4-amino-1-((4-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile(3.8 mg, 9.87 μmol, 13.89% yield, 98.77% purity), was obtained as awhite solid. LCMS (ESI+) m/z: 381.0.

1H NMR (400 MHz, DMSO-d₆): δ 11.66 (br, 1H), 8.57 (d, J=7.6 Hz, 1H),7.81 (d, J=7.6 Hz, 1H), 7.53 (t, J=7.8 Hz, 1H), 7.23 (s, 1H), 6.64 (s,1H), 6.48 (s, 1H), 5.79 (s, 2H), 4.17 (t, J=8.4 Hz, 2H), 3.07 (t, J=8.4Hz, 2H), 2.59 (s, 3H).

EXAMPLE 34: To a solution of4-amino-1-((4-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile(20 mg, 52.57 μmol, 1 eq) in DMF (2 mL) was added pyridine (12.48 mg,157.72 μmol, 12.73 μL, 3 eq) and MeI (74.62 mg, 525.74 μmol, 32.73 μL,10 eq). Then the mixture was stirred at 25° C. for 12 hr. The reactionmixture was concentrated under reduced pressure to give a residue. Theresidue was purified by prep-HPLC (column: Waters Xbridge BEH C18 100×25mm×5 m; mobile phase: [water (10 mM NH₄HCO₃)-ACN]; B %: 30%-60%, 8 min).4-(Dimethylamino)-1-((4-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile(2.1 mg, 5.09 μmol, 9.68% yield, 99% purity) was obtained as a whitesolid. LCMS (ESI+) m/z: 409.0.

¹H NMR (400 MHz, DMSO-d₆): δ 11.68 (br, 1H), 8.59 (d, J=7.6 Hz 1H), 7.86(d, J=7.6 Hz, 1H), 7.53 (t, J=8 Hz, 1H), 7.24 (s, 1H), 6.93 (s, 1H),6.78 (s, 1H), 4.17 (t, J=8.4 Hz, 2H), −3.05 (2H, mixed with solventpeak), 2.87 (s, 6H), 2.61 (s, 3H).

Examples 35, 36 & 37

N-(1-((4-chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)-6-cyanoindolin-4-yl)acetamide

4-amino-1-((4-chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile

1-((4-chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)-4-(dimethylamino)indoline-6-carbonitrile

Compound 27-2: To a solution of 4-chloroisoquinoline 27-1 (40 g, 244.49mmol, 1 eq) in H₂SO₄ (110 mL) was added dropwise a solution of KNO₃(32.13 g, 317.84 mmol, 1.3 eq) in H₂SO₄ (140 mL) slowly at −5° C. Themixture was stirred at 0° C. for 1 h, and then stirred at 15° C. for 12h. The mixture was poured into ice water (3 L) and then solid Na₂CO3 wasadded into the mixture until pH=8. A yellow solid was precipitated,filtered and the filter cake was washed with H₂O (2000 mL×3),concentrated the filter cake. The filter cake was dissolved in EtOAc,the EtOAc was filtered and the filter cake was concentrated underreduced pressure to give a residue. Crude Compound 27-2,4-chloro-5-nitroisoquinoline (43 g, 206.14 mmol, 84.31% yield), wasobtained as a yellow solid.

Compound 35-3: To a solution of 4-chloro-5-nitroisoquinoline 27-2 (43 g,206.14 mmol, 1 eq) in CHCl₃ (500 mL), m-CPBA (88.93 g, 412.27 mmol, 80%purity, 2 eq) was added in several portion at 0° C. The mixture stirredat 15° C. for 12 h. To the mixture was added sat. NaHCO₃(1 L) at 0° C.,the mixture was extracted with CHCl₃:MeOH=10:1 (500 mL×4) and then theorganic layers were washed with aq. 15% Na₂S₂SO₄ (500 mL×4), and thecombined organic layer was dried over Na₂SO₄, filtered and concentrated.The residue was purified by column chromatography (SiO₂, petroleumether:ethyl acetate=1:1 to 0:1). Compound 35-3,4-Chloro-5-nitroisoquinoline 2-oxide (36 g, 160.28 mmol, 77.76% yield)was obtained as a yellow solid.

Compound 35-4: To a solution of 4-chloro-5-nitroisoquinoline 2-oxide35-3 (36 g, 160.28 mmol, 1 eq) in DCM (500 mL), POCl₃ (49.15 g, 320.57mmol, 29.79 mL, 2 eq) was sloqly added at 0° C. The mixture was stirredat 60° C. for 12 h under N₂ atmosphere. The mixture was added into water(1000 mL) and extracted with DCM (500 mL×3). The organic layer waswashed with sat. NaHCO₃(800 mL×3) and brine (1500 mL), then dried overNa₂SO₄, filtered and concentrated. Crude Compound 35-4,1,4-dichloro-5-nitroisoquinoline (26 g, 106.98 mmol, 66.74% yield), wasobtained as a brown solid.

Compound 35-5: To a solution of 1,4-dichloro-5-nitroisoquinoline 35-4(25 g, 102.86 mmol, 1 eq) in MeOH (150 mL), NaOMe (24.45 g, 113.15 mmol,25% purity, 1.1 eq). The mixture was stirred at 80° C. for 1 h under N₂atmosphere, filtered and the filter cake was concentrated under reducedpressure to give Compound 36-5, 4-chloro-1-methoxy-5-nitroisoquinoline(16 g, 67.05 mmol, 65.19% yield), was obtained as a brown solid.

Compound 35-6: To a solution of 4-chloro-1-methoxy-5-nitroisoquinoline35-5 (15.9 g, 66.63 mmol, 1 eq) and NH₄Cl (35.64 g, 666.31 mmol, 10 eq)in MeOH (200 mL) and H₂O (40 mL), Zn (43.57 g, 666.31 mmol, 10 eq) wasslowly added at 0° C. The mixture was stirred at 15° C. for 2 hr,filtered and the filtrate was concentrated under reduced pressure togive a residue. The residue was added to H₂O (50 mL) and the mixture wasextracted with EtOAc (500 mL×4). The combined organic layers were washedwith brine (2000 mL×1), dried over Na₂SO₄, filtered and concentratedunder reduced pressure to give a residue. The residue was washed withpetroleum ether:ethyl acetate=30:1 (930 mL), filtered and the filtercake was dried under reduced pressure. Crude Compound 35-6,4-chloro-1-methoxyisoquinolin-5-amine (11.2 g, 53.68 mmol, 80.56%yield), was obtained as a white solid.

Preparation of Solution A: To a solution of4-chloro-1-methoxyisoquinolin-5-amine 35-6 (0.2 g, 958.57 μmol, 1 eq) inHCl (8 mL), a solution of NaNO₂ (99.21 mg, 1.44 mmol, 1.5 eq) in H₂O (1mL) at −5° C. was added. The mixture was stirred at −5° C. for 1 h.

Preparation of Solution B: SO₂ was bubbled into a solution of CuCl(23.72 mg, 239.64 mol, 5.73 μL, 0.25 eq) in AcOH (5 mL) and H₂O (1 mL)at 0° C. for 30 minutes at 15 psi.

Compound 35-7: Solution A was transferred into Solution B and themixture was stirred at 15° C. for 2 hr. The reaction mixture wasquenched by addition to ice water (20 mL) at 0° C., and then extractedwith DCM (30 mL×3). The combined organic layers were washed with brine(100 mL×2), dried over Na₂SO, filtered and concentrated under reducedpressure to give a residue. Compound 35-7,4-Chloro-1-methoxyisoquinoline-5-sulfonyl chloride (300 mg, crude), wasobtained as a yellow solid.

Compound 35-9: To a mixture of N-(6-cyanoindolin-4-yl)acetamide(Compound 30-4; 500 mg, 2.48 mmol, 1.3 eq) and pyridine (453.57 mg, 5.73mmol, 462.83 μL, 3 eq) was added dropwise a solution of4-chloro-1-methoxyisoquinoline-5-sulfonyl chloride 35-7 (1.27 g, 1.91mmol, 1 eq) in DCM (5 mL) at −5° C. The mixture was degassed and purgedwith N₂ 3 times, and the mixture was stirred at 15° C. for 12 hr underN₂ atmosphere. The reaction mixture was concentrated under reducedpressure to give a residue. Crude Compound 35-9,N-(1-((4-chloro-1-methoxyisoquinolin-5-yl)sulfonyl)-6-cyanoindolin-4-yl)acetamide(1.5 g, crude),was obtained as a red solid and used into next stepwithout further purification.

EXAMPLE 35: A mixture ofN-(1-((4-chloro-1-methoxyisoquinolin-5-yl)sulfonyl)-6-cyanoindolin-4-yl)acetamide35-9 (100 mg, 218.87 μmol, 1 eq) and TMSI (218.96 mg, 1.09 mmol, 148.96μL, 5 eq) in ACN (10 mL) was stirred at 80° C. for 2 hr under N₂atmosphere. The reaction mixture was quenched by addition to H₂O (20 mL)at 0° C., and then extracted with EtOAc (40 mL×2). The combined organiclayers were washed with brine (80 mL), dried over Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by prep-TLC (SiO₂, DCM: MeOH=10:1). Then the residue wasre-purified by prep-HPLC (column: Waters Xbridge BEH C18 100×25 mm×5 m;mobile phase: [water (10 mM NH₄HCO₃)-ACN]; B %: 20%-50%, 8 min). EXAMPLE35,N-(1-((4-Chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)-6-cyanoindolin-4-yl)acetamide(1.9 mg, 4.08 μmol, 1.86% yield, 95.1% purity), was obtained as a whitesolid. LCMS (ESI+) m/z: 443.0.

¹H NMR (400 MHz, DMSO-d₆): δ 9.73 (s, 1H), 8.57 (d, J=7.6 Hz, 1H), 8.01(d, J=8 Hz, 1H), 7.91 (s, 1H), 7.66 (s, 1H), 7.63 (t, J=7.6 Hz, 1H),6.98 (s, 1H), 4.19 (t, J=8.4 Hz, 2H), −3.20 (2H, mixed with solventpeak), 2.14 (s, 3H).

EXAMPLE 36: A solution ofN-(1-((4-chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)-6-cyanoindolin-4-yl)acetamideEXAMPLE 35 (16 mg, 36.13 μmol, 1 eq) in conc. HCl (3 mL) and EtOH (3 mL)was stirred at 50° C. for 5 hr. To the mixture was added saturatedNa₂CO₃ solution until pH=8 at 0° C., and the mixture was extracted withEtOAc (40 mL×5). The combined organic layers were washed with brine (200mL), dried over Na₂SO₄, filtered and concentrated under reduced pressureto give a residue. The residue was purified by prep-HPLC (column: WatersXbridge BEH C18 100×25 mm×5 μm; mobile phase: [water (10 mMNH₄HCO₃)-ACN]; B %: 20%-40%, 8 min). EXAMPLE 36,4-amino-1-((4-chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile(1.6 mg, 97.6% purity), was obtained as a white solid. LCMS (ESI+) m/z:400.9.

¹H NMR (400 MHz, DMSO-d₆): δ 12.04 (br, 1H), 8.56 (d, J=8 Hz, 1H), 7.92(d, J=7.6 Hz, 1H), 7.65˜7.61 (m, 2H), 6.63 (s, 1H), 6.45 (s, 1H), 5.77(s, 2H), 4.15 (t, J=8.4 Hz, 2H), 3.05 (t, J=8.4 Hz, 2H).

EXAMPLE 37: To a solution of4-amino-1-((4-chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrileEXAMPLE 36 (20 mg, 49.90 μmol, 1 eq) in DMF (2.5 mL) was added pyridine(39.47 mg, 498.95 μmol, 40.27 μL, 10 eq) and MeI (141.64 mg, 997.91 mol,62.12 μL, 20 eq). The mixture was stirred at 25° C. for 12 hr. Themixture was concentrated under reduced pressure to give a residue. Theresidue was purified by prep-HPLC (column: Waters Xbridge BEH C18 100×25mm×5 μm; mobile phase: [water (10 mM NH₄HCO₃)-ACN]; B %: 30%-60%, 8min). EXAMPLE 37,1-((4-Chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)-4-(dimethylamino)indoline-6-carbonitrile(3.1 mg, 7.08 tmol, 14.20% yield, 98% purity), was obtained as a whitesolid. LCMS (ESI+) m/z: 429.1.

¹H NMR (400 MHz, DMSO-d₆): δ 12.05 (br, 1H), 8.55 (d, J=8 Hz, 1H), 7.94(d, J=8 Hz, 1H), 7.65 (s, 1H), 7.63 (t, J=8 Hz, 1H), 6.9 2 (s, 1H), 6.74(s, 1H), 4.13 (t, J=8.4 Hz, 2H), ˜3.33 (2H, mixed with solvent peak),2.85 (s, 6H).

Example 38

1-((1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)-2,3-dihydro-1H-pyrrolo[3,2-b]pyridine-6-carbonitrile

EXAMPLE 38: To a mixture of2,3-dihydro-1H-pyrrolo[3,2-b]pyridine-6-carbonitrile 38A (21.45 mg,147.74 μmol, 1.2 eq) and pyridine (29.22 mg, 369.36 μmol, 29.81 μL, 3eq), was added dropwise a solution of 1-oxo-2H-isoquinoline-5-sulfonylchloride 1-4 (30 mg, 123.12 μmol, 1 eq) in DCM (3 mL) at −5° C. Themixture was degassed and purged with N₂ 3 times, and the mixture wasstirred at 25° C. for 12 hr under N₂ atmosphere. The reaction mixturewas concentrated under reduced pressure to give a residue. The crudeproduct was taken up in MeOH (2 mL) at 25° C. for 10 min, filteredconcentrated under reduced pressure to give a residue. The residue waspurified by prep-HPLC (column: Welch Ultimate AQ-C18 150×30 mm×5 M;mobile phase: [water(0.1% TFA)-ACN];B %: 35%-65%,12 min). EXAMPLE 38,1-[(1-oxo-2H-isoquinolin-5-yl)sulfonyl]-2,3-dihydropyrrolo[3,2-b]pyridine-6-carbonitrile(15 mg, 41.51 μmol, 33.71% yield, 97.5% purity), was obtained as a whitesolid. LCMS (ESI+) m/z: 353.0.

¹H NMR (400 MHz, DMSO-d₆): δ 11.71 (br, 1H), 8.59-8.52 (m, 2H), 8.46 (d,J=7.6 Hz, 1H), 7.90 (d, J=2 Hz, 1H), 7.64 (t, J=7.6 Hz, 1H), 7.39 (t,J=7.6 Hz, 1H), 7.11 (d, J=7.6 Hz, 1H), 4.08 (t, J=8.4 Hz, 2H), 3.22 (t,J=8.8 Hz, 2H).

Example 39

1-((4-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)-2,3-dihydro-1H-pyrrolo[3,2-b]pyridine-6-carbonitrile

Compound 39-2: To a mixture of 2,3-dihydro-1H-pyrrolo[3,2-b]pyridine-6-carbonitrile 38A (19.23 mg, 132.49μmol, 1.2 eq) and pyridine (26.20 mg, 331.22 μmol, 26.73 μL, 3 eq) wasadded dropwise a solution of 1-methoxy-4-methyl-isoquinoline-5-sulfonylchloride 32-8 (30 mg, 110.41 μmol, 1 eq) in DCM (2 mL) at −5° C. Themixture was degassed and purged with N₂ 3 times, and the mixture wasstirred at 25° C. for 12 hr under N₂ atmosphere. The reaction mixturewas concentrated under reduced pressure to remove DCM (2 mL). Theresidue was purified by prep-TLC (SiO₂, petroleum ether:ethylacetate=1:1). Compound 39-21-[(1-methoxy-4-methyl-5-isoquinolyl)sulfonyl]-2,3-dihydropyrrolo[3,2-b]pyridine-6-carbonitrile(15 mg, 39.43 μmol, 35.71% yield), was obtained as a white solid.

EXAMPLE 39: A mixture of1-[(1-methoxy-4-methyl-5-isoquinolyl)sulfonyl]-2,3-dihydropyrrolo[3,2-b]pyridine-6-carbonitrile39-2 (15 mg, 39.43 μmol, 1 eq) and TMSI (31.56 mg, 157.72 μmol, 21.47μL, 4 eq) in ACN (2 mL) was stirred at 25° C. for 1 hr under N₂atmosphere. The reaction mixture was quenched by addition of H₂O (5 mL)at 0° C., and then extracted with EtOAc (5 mL×2). The combined organiclayers were washed with brine (5 mL), dried over Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by prep-HPLC (column: Waters Xbridge Prep OBD C18 150×40 mm×10μm; mobile phase: [water(0.04% NH3H2O+10 mM NH4HCO3)-ACN];B %: 25%-55%,8min).

EXAMPLE 39,1-[(4-methyl-1-oxo-2H-isoquinolin-5-yl)sulfonyl]-2,3-dihydropyrrolo[3,2-b]pyridine-6-carbonitrile(3.4 mg, 9.28 μmol, 23.53% yield, 100% purity), was obtained as a whitesolid. LCMS (ESI+) m/z: 367.0.

1H NMR (400 MHz, DMSO-d₆): δ 11.72 (br, 1H), 8.62˜8.57 (m, 2H), 8.02 (d,J=2.4 Hz, 1H), 7.59˜7.51 (m, 2H), 7.25 (s, 1H), 4.23 (t, J=8.8 Hz, 2H),3.49 (t, J=8.8 Hz, 2H), 2.59 (s, 3H).

Example 40

1-((4-chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)-2,3-dihydro-1H-pyrrolo[3,2-b]pyridine-6-carbonitrile

Compound 40-2: To a mixture of2,3-dihydro-1H-pyrrolo[3,2-b]pyridine-6-carbonitrile (17.89 mg, 123.23μmol, 1.2 eq) 38A and pyridine (24.37 mg, 308.07 μmol, 24.87 μL, 3 eq)was added dropwise a solution of4-chloro-1-methoxy-isoquinoline-5-sulfonyl chloride 35-7 (30 mg, 102.69μmol, 1 eq) in DCM (2 mL) at −5° C. The mixture was degassed and purgedwith N₂ 3 times, and the mixture was stirred at 25° C. for 12 hr underN₂ atmosphere. LC-MS showed ˜50% of desired compound was detected. Thereaction mixture was concentrated under reduced pressure to remove DCM(2 mL). The residue was purified by prep-TLC (SiO₂, PE: EA=1:1).Compound 40-2, 1-[(4-chloro-1-methoxy-5-isoquinolyl)sulfonyl]-2,3-dihydropyrrolo[3,2-b]pyridine-6-carbonitrile (30 mg, 74.84μmol, 72.88% yield), was obtained as a white solid.

EXAMPLE 40: To a mixture of1-[(4-chloro-1-methoxy-5-isoquinolyl)sulfonyl]-2,3-dihydropyrrolo[3,2-b]pyridine-6-carbonitrile40-2 (30 mg, 74.84 μmol, 1 eq) and TMSI (59.90 mg, 299.37 μmol, 40.75μL, 4 eq) in acetonitrile (2 mL) was stirred at 25° C. for 1 hr under N₂atmosphere. The reaction mixture was quenched by addition of H₂O (4 mL)at 0° C., and then extracted with EtOAc (4 mL×2). The combined organiclayers were washed with brine (8 mL), dried over Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by prep-HPLC (column: Welch Ultimate AQ-C18 150×30 mm×5 μm;mobile phase: [water(0.1% TFA)-ACN];B %: 15%-45%,12 min). EXAMPLE 40,1-[(4-chloro-1-oxo-2H-isoquinolin-5-yl)sulfonyl]-2,3-dihydropyrrolo[3,2-b]pyridine-6-carbonitrile (7 mg, 17.55μmol, 23.45% yield, 97% purity), was obtained as a yellow solid. LCMS(ESI+) m/z: 386.9.

Example 41

1-(4-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)-4-(methylamino)indoline-6-carbonitrile

EXAMPLE 41: A mixture of4-amino-1-[(4-methyl-1-oxo-2H-isoquinolin-5-yl)sulfonyl]indoline-6-carbonitrile(70 mg, 184 μmol, 1 eq), (HCHO)_(n) (11 mg, 368.0 μmol, 2 eq), NaBH₃CN(23 mg, 368.0 μmol, 2 eq), glacial acetic acid (22 mg, 368.01 μmol, 21μL, 2 eq) in DMF (1 mL) was stirred at 60° C. for 12 hr. under N₂atmosphere. The reaction mixture was added saturated NaHCO₃(10 mL) andextracted with ethyl acetate (10 mL×3). The combined organic phase wasdried with anhydrous Na₂SO₄, filtered and concentrated under reducedpressure. The residue was purified by prep-HPLC (column: Phenomenex lunaC18 80×40 mm×3 μm; mobile phase: [water (0.04% HCl)-ACN]; B %: 37%-57%,7 min) to give4-(methylamino)-1-[(4-methyl-1-oxo-2H-isoquinolin-5-yl)sulfonyl]indoline-6-carbonitrileEXAMPLE 41 (5 mg, 13.2 μmol, 7.2% yield). Physical appearance: yellowsolid. LCMS (ESI+) m/z: 395.0. ¹H NMR (400 MHz, DMSO-d₆): δ 11.71 (br,1H), 8.58 (d, J=8 Hz, 1H), 7.8 (d, J=8 Hz, 1H), 7.53 (t, J=8 Hz, 1H),7.24 (d, J=5.2 Hz, 1H), 6.61 (s, 1H), 6.57 (s, 1H), 4.20 (t, J=8.4 Hz,2H), 3.09 (t, J=8.8 Hz, 2H), 2.79 (s, 3H), 2.60 (s, 3H)

Example 42

1-((4-chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)-4-(methylamino)indoline-6-carbonitrile

EXAMPLE 42: A mixture of4-amino-1-[(4-chloro-1-oxo-2H-isoquinolin-5-yl)sulfonyl]indoline-6-carbonitrileExample 36 (80 mg, 200 μmol, 1 eq), (HCHO)_(n) (12 mg, 400 μmol, 2 eq),NaBH₃CN (25 mg, 400 μmol, 2 eq), glacial acetic acid (24 mg, 400 μmol,23 μL, 2 eq) in DMF (1 mL) was stirred at 60° C. for 12 hr. The reactionmixture was added to saturated NaHCO₃(10 mL) and extracted with ethylacetate (10 mL×3). The combined organic phase was dried with anhydrousNa₂SO₄, filtered and concentrated under reduced pressure. The residuewas purified by prep-HPLC (column: Xtimate C18 100×30 mm×3 μm; mobilephase: [water (0.04% HCl)-ACN]; B %: 29%-49%, 9 min) to get1-[(4-chloro-1-oxo-2H-isoquinolin-5-yl)sulfonyl]-4-(methylamino)indoline-6-carbonitrile,EXAMPLE 42 (18 mg, 42 μmol, 21% yield). Physical appearance: yellowsolid. LCMS (ESI+) m/z: 415.0. ¹H NMR (400 MHz, DMSO-d₆): δ 12.09 (br,1H), 8.54 (d, J=8 Hz, 1H), 7.89 (d, J=8 Hz, 1H), 7.63-7.59 (m, 2H), 6.58(s, 1H), 6.53 (s, 1H), 4.15 (t, J=8.8 Hz, 2H), 3.05 (t, J=8.8 Hz, 2H),2.76 (s, 3H)

Example 43

4-(ethylamino)-1-((4-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile

EXAMPLE 43: A mixture of4-amino-1-[(4-methyl-1-oxo-2H-isoquinolin-5-yl)sulfonyl]indoline-6-carbonitrile,Example 33 (50 mg, 131 μmol, 1 eq), acetaldehyde (5 M in THF, 395 μL, 15eq), NaBH₃CN (25 mg, 394 μmol, 3 eq), glacial acetic acid (24 mg, 395μmol, 23 μL, 3 eq) in DMF (1 mL) was stirred at 60° C. for 12 hr. in asealed tube. The reaction mixture was added to sat. NaHCO₃(40 mL) andextracted with ethyl acetate (40 mL×3). The combined organic phase wasdried with anhydrous Na₂SO₄, filtered and concentrated under reducedpressure. The residue was purified by prep-HPLC (column: Waters XbridgePrep OBD C18 150×40 mm×10 μm; mobile phase: [water (10 mM NH₄HCO₃)-ACN];B %: 35%-55%, 8 min) to give4-(ethylamino)-1-[(4-methyl-1-oxo-2H-isoquinolin-5-yl)sulfonyl]indoline-6-carbonitrileIND-12202019-2 (4.3 mg, 8% yield). Physical appearance: yellow solid.LCMS (ESI+) m/z: 409.1. ¹H NMR (400 MHz, DMSO-d₆): δ 8.57 (d, J=7.6 Hz,1H), 7.81 (d, J=7.6 Hz, 1H), 7.53 (t, J=8 Hz, 1H), 7.23 (s, 1H), 6.65(s, 1H), 6.54 (s, 1H), 5.84 (t, J=5.6 Hz, 1H), 4.18 (t, J=8.4 Hz, 2H),3.22-3.12 (m, 2H), 3.09 (t, J=8.4 Hz, 2H), 2.59 (s, 3H), 1.19 (t, J=8.8Hz, 3H)

Example 44

1-((4-chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)-4-(ethylamino)indoline-6-carbonitrile

EXAMPLE 44: A mixture of4-amino-1-[(4-chloro-1-oxo-2H-isoquinolin-5-yl)sulfonyl]indoline-6-carbonitrileExample 36 (80 mg, 200 μmol, 1 eq), acetaldehyde (5 M, 600 μL, 15 eq),NaBH₃CN (38 mg, 600 μmol, 3 eq), glacial acetic acid (36 mg, 600 μmol,34.24 L, 3 eq) in DMF (1 mL) was stirred at 60° C. for 12 hr. in asealed tube. The reaction mixture (combined with another batch of 20 mgscale) was added sat. NaHCO₃(20 mL) and extracted with ethyl acetate (20mL×3). The combined organic phase was dried with anhydrous Na₂SO₄,filtered and concentrated under reduced pressure. The residue waspurified by prep-HPLC (column: Waters Xbridge Prep OBD C18 150×40 mm×10um; mobile phase: [water (10 mM NH₄HCO₃)-ACN]; B %: 35%-55%, 8 min) toget1-[(4-chloro-1-oxo-2H-isoquinolin-5-yl)sulfonyl]-4-(ethylamino)indoline-6-carbonitrile,EXAMPLE 44 (17 mg, 16% yield). Physical appearance: yellow solid. LCMS(ESI+) m/z: 429.0. ¹H NMR (400 MHz, DMSO-d₆): δ 11.90 (br, 1H), 8.55 (d,J=8 Hz, 1H), 7.91 (d, J=8 Hz, 1H), 7.64-7.60 (m, 2H), 6.63 (s, 1H), 6.51(s, 1H), 5.82 (t, J=5.6 Hz, 1H), 4.15 (t, J=8.8 Hz, 2H), 3.21-3.12 (m,2H), 3.09 (t, J=8.8 Hz, 2H), 1.18 (t, J=7.2 Hz, 3H)

Example 45

4-((2-((1R,4R)-2-oxa-5-azabicyclo[2.2.1]heptan-5-yl)ethyl)amino)-1-((4-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile

45-2: To a mixture of (1R,4R)-2-oxa-5-azabicyclo[2.2.1]heptane, 45-1 (1g, 10.1 mmol, 1 eq) and 2-bromo-1,1-diethoxy-ethane (2.6 g, 13.11 mmol,2 mL, 1.3 eq) in DMF (10 mL) was added K₂CO₃ (2.8 g, 20.2 mmol, 2 eq)and KI (167 mg, 1.01 mmol, 0.1 eq) under N₂. The mixture was stirred at100° C. for 12 hours. The reaction mixture was filtered, and thefiltrate was concentrated under reduced pressure. The residue waspurified by column chromatography (SiO₂, Petroleum ether: Ethylacetate=20:1 to 3:1) to give(1S,4S)-5-(2,2-diethoxyethyl)-2-oxa-5-azabicyclo[2.2.1]heptane 45-2(1.73 g, 8.2 mmol, 81% yield). Physical appearance: yellow solid. LCMS(ESI+) m/z: 216.0. ¹H NMR (400 MHz, CDCl₃): δ4.47 (t, J=5.2 Hz, 1H),4.29 (s, 1H), 3.94 (d, J 7.6 Hz, 1H), 3.77-3.44 (m, 6H), 2.94 (dd, J=10Hz, J=1.6 Hz, 1H), 2.74-2.63 (m, 2H), 2.54 (d, J=10 Hz, 1H), 1.77 (dd,J=9.6 Hz, J=2.0 Hz, 1H), 1.64 (d, J=9.6 Hz, 1H), 1.21-1.09 (m, 6H).

Example 45: To a mixture of4-amino-1-[(4-methyl-1-oxo-2H-isoquinolin-5-yl)sulfonyl]indoline-6-carbonitrileExample 33 (80 mg, 210 μmol, 1 eq) and(1R,4R)-5-(2,2-diethoxyethyl)-2-oxa-5-azabicyclo[2.2.1]heptane 45-2 (136mg, 631 μmol, 3 eq) in DCM (4 mL) was added TFA (336 mg, 3 mmol, 218 μL,14 eq) and Et₃SiH (245 mg, 2.1 mmol, 336 μL, 10 eq). The mixture wasstirred at 60° C. for 12 hr. The reaction mixture was added saturatedNaHCO₃(15 mL) and extracted with DCM (15 mL×3). The combined organicphase was dried with anhydrous Na₂SO₄, filtered and concentrated underreduced pressure. The residue was purified by prep-HPLC (column: XtimateC18 100×30 mm×3 μm; mobile phase: [water (0.04% HCl)-ACN]; B %: 14%-44%,9 min) to give1-[(4-methyl-1-oxo-2H-isoquinolin-5-yl)sulfonyl]-4-[2-[(1R,4R)-2-oxa-5-azabicyclo[2.2.1]heptan-5-yl]ethylamino]indoline-6-carbonitrile,EXAMPLE 45 (5.14 mg, 9.5 μmol, 4.54% yield, 94% purity) as hydrochloridesalt. Physical appearance: yellow solid. LCMS (ESI+) m/z: 506.1. ¹H NMR(400 MHz, DMSO-d₆): δ 11.70 (d, J=5.6 Hz, 1H), 10.96 (m, ˜0.5H), 10.44(m, ˜0.5H), 8.59 (d, J=8 Hz, 1H), 7.82 (d, J=8 Hz, 1H), 7.53 (t, J=8 Hz,1H), 7.24 (d, J=5.6 Hz, 1H), 6.85-6.80 (m, 1H), 6.62 (s, 1H), 6.25-6.15(m, 1H), 4.70-4.58 (m, 2H), 4.24-4.15 (m, 3H), 3.76-3.72 (m, 1H),3.62-3.51 (m, 3H), 3.18 (t, J=8.8 Hz, 3H), 3.07-3.04 (m, 1H), 2.60 (s,3H), 2.34-2.25 (m, 1H), 2.19-2.13 (m, 1H), 2.03-1.96 (m, 1H)

Example 46

4-((2-((1S,4S)-2-oxa-5-azabicyclo[2.2.1]heptan-5-yl)ethyl)amino)-1-((4-chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile

Example 46: To a mixture of4-amino-1-[(4-chloro-1-oxo-2H-isoquinolin-5-yl)sulfonyl]indoline-6-carbonitrileExample 36 (80 mg, 210 μmol, 1 eq) and(1R,4R)-5-(2,2-diethoxyethyl)-2-oxa-5-azabicyclo[2.2.1]heptane 45-2 (136mg, 631 μmol, 3 eq) in DCM (4 mL) was added TFA (336 mg, 2.94 mmol, 218μL, 14 eq) and Et₃SiH (245 mg, 2.1 mmol, 336 μL, 10 eq). The mixture wasstirred at 60° C. for 12 hr. The reaction mixture was added saturatedNaHCO₃(15 mL) and extracted with DCM (15 mL×3). The combined organicphase was dried with anhydrous Na₂SO₄, filtered and concentrated underreduced pressure. The residue was purified by prep-HPLC (column: XtimateC18 100×30 mm×3 m; mobile phase: [water (0.04% HCl)-ACN]; B %: 14%-44%,9 min) to give1-[(4-methyl-1-oxo-2H-isoquinolin-5-yl)sulfonyl]-4-[2-[(1R,4R)-2-oxa-5-azabicyclo[2.2.1]heptan-5-yl]ethylamino]indoline-6-carbonitrile,EXAMPLE 46 (3.5 mg, 6.5 μmol, 3% yield, 91% purity) as hydrochloridesalt. Physical appearance: yellow solid. LCMS (ESI+) m/z: 506.1.

Example 47

4-((2-((1S,4S)-2-oxa-5-azabicyclo[2.2.1]heptan-5-yl)ethyl)amino)-1-((4-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile

47-2: A mixture of (1S,4S)-2-oxa-5-azabicyclo[2.2.1]heptane 47-1 (2 g,20.18 mmol, eq), 2-bromo-1,1-diethoxy-ethane (5.17 g, 26.23 mmol, 3.95mL, 1.3 eq), K₂CO₃ (5.58 g, 40.35 mmol, 2 eq), KI (335 mg, 2.02 mmol,0.1 eq) in DMF (20 mL) was degassed and purged with N₂ for 3 times, andthen the mixture was stirred at 100° C. for 12 hr. under N₂ atmosphere.The reaction mixture was filtered, and the filtrate was concentratedunder reduced pressure. The residue was purified by columnchromatography (SiO₂, Petroleum ether: Ethyl acetate=20:1 to 3:1) togive (1S,4S)-5-(2,2-diethoxyethyl)-2-oxa-5-azabicyclo[2.2.1]heptane 47-2(3.5 g, 16.26 mmol, 80.58% yield). Physical appearance: yellow oil. LCMS(ESI+) m/z: 216.0. ¹H NMR (400 MHz, CDCl₃): δ4.53 (t, J=5.2 Hz, 1H),4.29 (s, 1H), 4.01 (d, J=8.0 Hz, 1H), 3.72-3.52 (m, 6H), 3.00 (dd, J=10Hz, J=1.6 Hz, 1H), 2.80-2.69 (m, 2H), 2.60 (d, J=10 Hz, 1H), 1.84 (dd,J=9.6 Hz, J=2.0 Hz, 1H), 1.70 (d, J=9.6 Hz, 1H), 1.21 (t, J=8.2 Hz, 6H).

Example 47: To a mixture of4-amino-1-[(4-methyl-1-oxo-2H-isoquinolin-5-yl)sulfonyl]indoline-6-carbonitrileExample 33 (100 mg, 263 μmol, 1 eq),(1S,4S)-5-(2,2-diethoxyethyl)-2-oxa-5-azabicyclo[2.2.1]heptane 47-2 (170mg, 790 μmol, 3 eq) in DCM (5 mL) was added TFA (420 mg, 3.68 mmol, 273μL, 14 eq) and Et₃SiH (306 mg, 2.63 mmol, 420 μL, 10 eq), then themixture was stirred at 60° C. for 12 hr. in a sealed tube. The reactionmixture was added sat. NaHCO₃(15 mL) and extracted with DCM (15 mL×3).The combined organic phase was dried with anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure. The residue was purified byprep-HPLC (column: Xtimate C18 100×30 mm×3 μm; mobile phase: [water(0.04% HCl)-ACN]; B %: 14%-34%, 9 min) to get1-[(4-methyl-1-oxo-2H-isoquinolin-5-yl)sulfonyl]-4-[2-[(1S,4S)-2-oxa-5-azabicyclo[2.2.1]heptan-5-yl]ethylamino]indoline-6-carbonitrileEXAMPLE 47 (1.12 mg, 2.62 μmol, 1% yield). Physical appearance: yellowsolid. LCMS (ESI+) m/z: 506.2. ¹H NMR (400 MHz, DMSO-d₆): δ 11.70 (d,J=5.6 Hz, 1H), 10.98 (br, ˜0.5H), 10.45 (br, ˜0.5H), 8.58 (d, J=8 Hz,1H), 7.81 (d, J=7.6 Hz, 1H), 7.52 (t, J=8 Hz, 1H), 7.24 (d, J=6 Hz, 1H),6.83 (d, J=5.6 Hz, 1H), 6.61 (s, 1H), 6.25˜6.15 (m, 1H), 4.69-4.58 (m,2H), 4.23-4.14 (m, 3H), 3.77-3.72 (m, 1H), 3.60-3.50 (m, 3H), 3.17 (t,J=8.8 Hz, 3H), 3.07-3.03 (m, 1H), 2.60 (s, 3H), 2.36-2.28 (m, 1H),2.19-2.09 (m, 1H), 2.02-1.92 (m, 1H)

Example 48

4-((2-((1S,4S)-2-oxa-5-azabicyclo[2.2.1]heptan-5-yl)ethyl)amino)-1-((4-chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)indoline-6-carbonitrile

Preparation of EXAMPLE 48: To a mixture of4-amino-1-[(4-chloro-1-oxo-2H-isoquinolin-5-yl)sulfonyl]indoline-6-carbonitrileExample 36 (100 mg, 250 μmol, 1 eq),(1S,4S)-5-(2,2-diethoxyethyl)-2-oxa-5-azabicyclo[2.2.1]heptane 47-2 (161mg, 750 μmol, 3 eq) in DCM (5 mL) was added TFA (400 mg, 3.50 mmol,258.60 μL, 14 eq) and Et₃SiH (290 mg, 2.5 mmol, 400 μL, 10 eq), then themixture was stirred at 60° C. for 12 hr. in a sealed tube. The reactionmixture was added sat. NaHCO₃(15 mL) and extracted with DCM (15 mL×3).The combined organic phase was dried with anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure. The residue was purified byprep-HPLC (column: Xtimate C18 100×30 mm×3 μm; mobile phase: [water(0.04% HCl)-ACN]; B %: 14%-34%, 9 min) to get1-[(4-chloro-1-oxo-2H-isoquinolin-5-yl)sulfonyl]-4-[2-[(1S,4S)-2-oxa-5-azabicyclo[2.2.1]heptan-5-yl]ethylamino]indoline-6-carbonitrileEXAMPLE 48 (3.33 mg, 6.33 μmol, 2.5% yield). Physical appearance: yellowsolid. LCMS (ESI+) m/z: 526.1. ¹H NMR (400 MHz, DMSO-d₆): δ 12.12 (br,1H), 11.40 (br, ˜0.5H), 10.94 (br, ˜0.5H), 8.55 (d, J=8.8 Hz, 1H), 7.92(d, J=8 Hz, 1H), 7.65-7.59 (m, 2H), 6.82 (d, J=6.4 Hz, 1H), 6.59 (d, J=4Hz, 1H), 6.30-6.21 (m, 1H), 4.68-4.58 (m, 2H), 4.30-4.10 (m, 3H),3.73-3.69 (m, 1H), 3.60-3.50 (m, 3H), 3.16 (t, J=8 Hz, 3H), 3.07-3.04(m, 1H), 2.33-2.25 (m, 1H), 2.24-2.13 (m, 1H), 1.99-1.90 (m, 1H)

Example 49

1-((4-methyl-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)-4-(oxetan-3-ylamino)indoline-6-carbonitrile

EXAMPLE 49: A mixture of4-amino-1-[(4-methyl-1-oxo-2H-isoquinolin-5-yl)sulfonyl]indoline-6-carbonitrileExample 33 (100 mg, 263 μmol, 1 eq), oxetan-3-one (380 mg, 5.3 mmol, 20eq), NaBH₃CN (50 mg, 790 μmol, 3 eq), TFA (150 mg, 1.31 mmol, 97 μL, 5eq) and 4 Å molecular sieve (20 mg, 250 μmol) in i-PrOH (5 mL) and DMF(5 mL) was stirred at 60° C. for 12 hr. under N₂ atmosphere. Thereaction mixture was added sat. NaHCO₃(15 mL) and extracted with DCM (15mL×3). The combined organic phase was dried with anhydrous Na₂SO₄,filtered and concentrated under reduced pressure. The residue waspurified by prep-HPLC (column: Waters Xbridge Prep OBD C18 150×40 mm×10m; mobile phase: [water (10 mM NH₄HCO₃)-ACN]; B %: 25%-45%, 8 min) toget1-[(4-methyl-1-oxo-2H-isoquinolin-5-yl)sulfonyl]-4-(oxetan-3-ylamino)indoline-6-carbonitrileEXAMPLE 49 (21.1 mg, 48.3 μmol, 18% yield). Physical appearance: whitesolid. LCMS (ESI+) m/z: 437.0. ¹H NMR (400 MHz, DMSO-d₆): δ 8.57 (dd,J=8 Hz, J=1.6 Hz, 1H), 7.83 (dd, J=7.6 Hz, J=1.2 Hz, 1H), 7.53 (t, J=8Hz, 1H), 7.23 (s, 1H), 6.61 (s, 1H), 6.51 (d, J=6.4 Hz, 1H), 6.47 (s,1H), 4.88 (t, J=6.4 Hz, 2H), 4.72-4.65 (m, 1H), 4.54 (t, J=6 Hz, 2H),4.20 (t, J=8.8 Hz, 2H), 3.17 (t, J=8.8 Hz, 2H), 2.59 (s, 3H)

Example 50

1-((4-chloro-1-oxo-1,2-dihydroisoquinolin-5-yl)sulfonyl)-4-(oxetan-3-ylamino)indoline-6-carbonitrile

EXAMPLE 50: A mixture of4-amino-1-[(4-chloro-1-oxo-2H-isoquinolin-5-yl)sulfonyl]indoline-6-carbonitrileExample 36 (100 mg, 249.48 μmol, 1 eq), oxetan-3-one (360 mg, 5 mmol, 20eq), NaBH₃CN (47 mg, 750 μmol, 3 eq), TFA (142 mg, 1.25 mmol, 92 μL, 5eq) and 4 Å molecular sieve (20 mg, 250 μmol) in i-PrOH (5 mL) and DMF(5 mL) was stirred at 60° C. for 12 hr. under N₂ atmosphere. Thereaction mixture was added saturated NaHCO₃ solution (15 mL) andextracted with DCM (15 mL×3). The combined organic phase was dried withanhydrous Na₂SO₄, filtered and concentrated under reduced pressure. Theresidue was purified by prep-HPLC (column: Waters Xbridge Prep OBD C18150×40 mm×10 μm; mobile phase: [water (10 mM NH₄HCO₃)-ACN]; B %:25%-45%, 8 min) to get1-[(4-chloro-1-oxo-2H-isoquinolin-5-yl)sulfonyl]-4-(oxetan-3-ylamino)indoline-6-carbonitrileEXAMPLE 50 (8.3 mg, 18 μmol, 7.3% yield). Physical appearance: whitesolid. LCMS (ESI+) m/z: 457.0. ¹H NMR (400 MHz, DMSO-d₆): δ 8.57 (d, J=8Hz, 1H), 7.92 (d, J=6.8 Hz, 1H), 7.66 (s, 1H), 7.60 (t, J=8 Hz, 1H),6.58 (s, 1H), 6.50 (d, J=6.4 Hz, 1H), 6.45 (s, 1H), 4.87 (t, J=6.8 Hz,2H), 4.70-4.65 (m, 1H), 4.53 (t, J=6 Hz, 2H), 4.17 (t, J=8.8 Hz, 2H),3.14 (t, J=8.8 Hz, 2H)

Indoline Reagents Indonline 2

6-Chloroindoline

INDOLINE 2: To a mixture of 6-chloro-1H-indole (5 g, 32.98 mmol) in AcOH(80 mL) was added NaBH₃CN (6.84 g, 108 mmol) at 0° C., and then themixture was stirred at 25° C. for 12 h under an N₂ atmosphere. Thereaction mixture was quenched by the addition of ice water (20 mL) at 0°C. and then saturated aq. NaOH was added to the mixture to adjust the pHto ˜8. The mixture was extracted with EtOAc (250 mL×2). The combinedorganic layers were dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by column chromatography (SiO₂, petroleum ether:ethylacetate=20:1 to 5:1). The title compound, 6-chloroindoline INDOLINE 2 (3g, 19.5 mmol, 59.2% yield), was obtained as green oil.

Indonline 4

Indoline-6-carbonitrile

INDOLINE 4: To a mixture of 1H-indole-6-carbonitrile (5 g, 35.1 mmol) inAcOH (80 mL) was added NaBH3CN (7.29 g, 116.0 mmol) at 0° C., and thenthe mixture was stirred at 25° C. for 12 h under an N₂ atmosphere. Thereaction mixture was quenched by addition of ice water (20 mL) at 0° C.Saturated aq. NaOH was added to the mixture to adjust pH to −8. Themixture was extracted with EtOAc (300 mL×2). The combined organic layerswere dried over anhydrous Na₂SO₄, filtered and concentrated underreduced pressure to give a residue. The residue was purified by columnchromatography (SiO₂, petroleum ether:ethyl acetate=10:1 to 5:1). Theresidue was purified by prep-HPLC (column: Xtimate C18 10μ×250 mm×50 mm;mobile phase:[A, water(10 mM NH₄HCO₃)—B, ACN]; B %: 20%-40%, 25 min).The title compound, indoline-6-carbonitrile INDOLINE 4 (1.9 g, 13.1mmol, 37.4% yield), was obtained as a white solid.

Indonline 8

1-(Indolin-4-yl)ethan-1-ol

Step 1: To a solution of 1H-indole-4-carbonitrile (5 g, 35.1 mmol) inTHF (20 mL) was added MeLi (1.8 M, 41.9 mL). The mixture was stirred at80° C. for 8 h under an N₂ atmosphere. The reaction mixture was quenchedby addition of H₂O (100 mL) at 0° C. The reaction mixture was extractedwith DCM (400 mL×2). The combined organic layers were dried overanhydrous Na₂SO₄, filtered and concentrated under reduced pressure togive a residue. The residue was purified by column chromatography (SiO₂,petroleum ether:ethyl acetate=20:1 to 1:1). The title compound,1-(1H-indol-4-yl) ethanone (2.5 g, 15.7 mmol, 44.6% yield), was obtainedas a yellow solid.

Step 2: To a solution of 1-(1H-indol-4-yl)ethanone (2.4 g, 15.0 mmol) inTHF (20 mL) was added NaBH₄ (2.28 g, 60.3 mmol) at 0° C. The mixture wasstirred at 25° C. for 12 h under N₂ atmosphere. The reaction mixture wasquenched by addition of H₂O (100 mL) at 0° C. The reaction mixture wasextracted with DCM (250 mL×2). The combined organic layers were driedover anhydrous Na₂SO₄, filtered and concentrated under reduced pressureto give a residue. The residue was purified by column chromatography(SiO₂, petroleum ether:ethyl acetate=1:1). 1-(1H-Indol-4-yl)ethanol (1g, 6.20 mmol, 41.1% yield) was obtained as a white solid.

INDOLINE 8, Step 3: To a mixture of 1-(1H-indol-4-yl)ethanol (1 g, 6.20mmol) in AcOH (10 mL) was added NaBH₃CN (1.29 g, 20.4 mmol) at 0° C.,and then the mixture was stirred at 25° C. for 5 h under an N₂atmosphere. The reaction mixture was quenched by addition of ice water(15 mL) at 0° C., and then saturated aq. NaOH was added to the mixtureto adjust the pH to −8. The reaction mixture was extracted with DCM (100mL×2). The combined organic layers were dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to give a residue. Theresidue was purified by column chromatography (SiO₂, petroleumether:ethyl acetate=10:1 to 1:1). 1-Indolin-4-ylethanol INDOLINE 8 (250mg, 1.53 mmol, 24.6% yield) was obtained as a white solid.

Indonline 10

4,6-Dichloroindoline

INDOLINE 10: To a mixture of 4,6-dichloro-1H-indole (0.5 g, 2.69 mmol)in AcOH (5 mL) was added NaBH₃CN (557 mg, 8.87 mmol) at 0° C., and thenthe mixture was stirred at 25° C. for 12 h under an N₂ atmosphere. Thereaction mixture was quenched by the addition of ice water (20 mL) at 0°C., and then saturated aq. NaOH was added to the mixture to adjust thepH to −8. The reaction mixture was extracted with DCM (100 mL×2). Thecombined organic layers were dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by column chromatography (SiO₂, petroleum ether:ethylacetate=30:1 to 5:1). 4,6-Dichloroindoline INDOLINE 10 (0.1 g, 531.7μmol, 19.7% yield) was obtained as yellow oil.

Indonline 13

4-Cyclopropylindoline-6-carbonitrile

Step 1: A mixture of 4-bromo-1H-indole-6-carbonitrile (0.1 g, 452.3μmol), cyclopropylboronic acid (77.7 mg, 904.7 μmol), Cs₂CO₃ (294.7 mg,904.7 μmol), Pd(dppf)Cl₂ (33.1 mg, 45.2 μmol) in dioxane (5 mL) and H₂O(0.5 mL) was degassed and purged with N₂ 3 times. Then the mixture wasstirred at 110° C. for 12 h under an N₂ atmosphere. The reaction mixturewas diluted with H₂O (20 mL) and extracted with EtOAc (80 mL×2). Thecombined organic layers were dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by column chromatography (SiO₂, petroleum ether:ethylacetate=20:1 to 5:1). 4-Cyclopropy 1-1H-indole-6-carbonitrile (50 mg,274.3 μmol, 60.6% yield) was obtained as a yellow solid.

INDOLINE 13, Step 2: To a solution of4-cyclopropyl-1H-indole-6-carbonitrile (50 mg, 274.3 μmol) in AcOH (3mL) was added sodium; cyanoboranuide (56.90 mg, 905.4 μmol) at 0° C. Themixture was stirred at 25° C. for 12 h. The reaction mixture wasquenched by the addition of ice water (5 mL) at 0° C., and thensaturated aq. NaOH was added to the mixture to adjust pH to ˜8. Thereaction mixture was extracted with DCM (50 mL×2). The combined organiclayers were dried over anhydrous Na₂SO₄, filtered and concentrated underreduced pressure to give a residue. The residue was purified by prep-TLC(SiO₂, petroleum ether:ethyl acetate=5:1). Compound4-cyclopropylindoline-6-carbonitrile INDOLINE 13 (20 mg, 108.6 μmol,39.5% yield) was obtained as a yellow solid.

Indonline 16

4-Methylindoline-6-carbonitrile

Step 1: To a mixture of 4-bromo-1H-indole-6-carbonitrile (1 g, 4.52mmol) in dioxane (15 mL) and H₂O (1.5 mL) was added methylboronic acid(541.5 mg, 9.05 mmol), Cs₂CO₃ (2.95 g, 9.05 mmol) and Pd(dppf)Cl₂ (331.0mg, 452.3 μmol) in one portion. The mixture was heated to 110° C. andstirred for 12 h. The reaction mixture was diluted with H₂O (300 mL) andextracted with EtOAc (150 mL×2). The combined organic layers were washedwith brine (300 mL), dried over Na₂SO₄, filtered and concentrated underreduced pressure to give a residue. The residue was purified by columnchromatography (SiO₂, petroleum ether:ethyl acetate=1:0 to 0:1).4-Methyl-1H-indole-6-carbonitrile (600 mg, 3.34 mmol, 73.8% yield) wasobtained as a light yellow solid.

INDOLINE 16, Step 2: To a solution of 4-methyl-1H-indole-6-carbonitrile(600 mg, 3.84 mmol) in HOAc (15 mL) was added NaBH₃CN (1.45 g, 23.0mmol). The mixture was stirred at 25° C. for 12 h. The reaction mixturewas added saturated aq. Na₂CO₃ by pH to ˜8.

The mixture was diluted with H₂O (300 mL) and extracted with EtOAc (150mL×3). The combined organic layers were washed with brine (300 mL),dried over Na₂SO₄, filtered and concentrated under reduced pressure togive a residue. The residue was purified by column chromatography (SiO₂,petroleum ether:ethyl acetate=1:0 to 0:1).4-Methylindoline-6-carbonitrile (250 mg, 1.20 mmol, 31.2% yield) wasobtained as a light yellow solid.

Indonline 18

4-(Prop-1-en-2-yl)indoline

INDOLINE 18: A mixture of 4-bromoindoline (0.5 g, 2.52 mmol),2-isopropenyl-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (509.0 mg, 3.03mmol), Cs₂CO₃ (1.23 g, 3.79 mmol), Pd(dppf)Cl₂ (184.7 mg, 252.4 μmol) indioxane (10 mL) and H₂O (2 mL) was degassed and purged with N₂ 3 times,and then the mixture was stirred at 110° C. for 12 h under an N₂atmosphere. The reaction mixture was diluted with H₂O (20 mL) andextracted with DCM (60 mL×3). The combined organic layers were washedwith saturated brine (20 mL), dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by column chromatography (SiO₂, petroleum ether:ethylacetate=1:0 to 10:1). 4-Isopropenylindoline (60 mg, 376.8 μmol, 14.9%yield) was obtained as a yellow solid.

Indonline 20

4-Vinylindoline

INDOLINE 20: To a mixture of 4-bromoindoline (1 g, 5.05 mmol) and4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (933 mg, 6.06 mmol) indioxane (10 mL) and H₂O (2 mL) as added Pd(dppf)Cl₂ (369 mg, 504.9 μmol)and Cs₂CO₃ (2.47 g, 7.57 mmol) at 15° C. under N₂. The mixture washeated to 110° C. and stirred for 12 hours. The residue was diluted withH₂O (120 mL) and extracted with EtOAc (60 mL×2). The combined organiclayers were washed with brine 120 mL, dried over Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by column chromatography (SiO₂, petroleum ether:ethylacetate=1:0 to 10:1). 4-Vinylindoline INDOLINE 20 (400 mg, 2.45 mmol,48.6% yield) was obtained as a yellow oil.

Indonline 22

4-Cyclopropylindoline

INDOLINE 22: To a solution of 4-bromoindoline (0.5 g, 2.52 mmol) indioxane (10 mL) and H₂O (1 mL) was added cyclopropylboronic acid (433.6mg, 5.05 mmol), dicesium;carbonate (1.65 g, 5.05 mmol) andcyclopentyl(diphenyl)phosphane;dichloropalladium;iron (184.7 mg, 252.4μmol). The mixture was stirred at 110° C. for 12 h under an N₂atmosphere. The reaction mixture was diluted with H₂O 15 mL andextracted with EtOAc (40 mL×2). The combined organic layers were washedwith saturated brine 20 mL, dried over anhydrous Na₂SO₄, filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by column chromatography (SiO₂, petroleum ether:ethylacetate=1:0 to 10:1). 4-Cyclopropylindoline INDOLINE 22 (0.1 g, 628 mol,24.8% yield) was obtained as yellow oil.

Example 51

This Example illustrates that exemplary compounds of the presentinvention inhibit ROCK1 and/or ROCK2 enzymatic activity.

Dose-response curves for compounds of the present invention weredetermined using a fluorescence coupling enzymatic assay using purifiedC-terminal truncated human ROCK1 catalytic domain (amino acids 1-477 ofaccession number NP_005397.1) fused to GST (Carna BioSciences, Catalog#01-109) and purified C-terminal truncated human ROCK2 enzyme kinasedomain (amino acids 1-553 of accession number NP_004841.2) fused to GST(Carna BioSciences, Catalog #01-110). In addition, full lengthcGMP-dependent protein kinase 1 (PKG or PRKG1; amino acids 1-686 ofaccession number NP_006249.1) fused to GST (Carna BioSciences, Catalog#01-142), full length PKACa (PRKACA; amino acids 1-351 of accessionnumber NP_002721.1) fused to GST (Carna BioSciences, Catalog #01-127),and C-terminal truncated human AKT1 catalytic domain (amino acids104-480 of accession number NP_005154.1) fused to GST (CarnaBiosciences, Catalog #01-101) were used as a control to measure anynon-desired off-target kinase activity.

Briefly, compounds of the present invention were solubilized in DMSO anda series of 12, three-fold serial dilutions were made for each compoundin 2.5% DMSO. The initial starting concentration for the serialdilutions of each compound was 10 μM or 100 μM. Control samples lackingcompound, ROCK enzyme or ATP also were prepared and processed inparallel with compound test samples.

A 4 μL aliquot of each serial dilution of test compound diluted in H₂Owith 2.5% DMSO was added to a well containing 4 μL of 10 nM purifiedN-truncated ROCK1 (4 nM final concentration), 10 nM purified N-truncatedROCK2 enzyme (4 nM final concentration), 1.25 nM AKT1 (0.5 nM finalconcentration), 25 pM PKACA (10 pM final concentration) or 20 nM PKG (8nM final concentration), in reaction buffer containing 50 mM Hepes, pH7.5, 1 mM EGTA, 10 mM MgCl₂, 2 mM DTT, and 0.01% Tween 20 (finalconcentration). To initiate the enzymatic reaction, 2 μL of a solutionwas added to each well to provide a final concentration of 50 nMUlight-CREBTide peptide substrate (Perkin Elmer, Catalog # TRF0107) and4 μM ATP for ROCK1 and AKT1 assays, 8 μM ATP for ROCK2 assays and 30 μMATP for PKG assays, or 1 μM ATP for PKACA assays. The reaction mixtureswere incubated at room temperature for one hour. After one hour, thereaction was stopped by the addition of a 10 μL solution of anEuropium-labelled phospho-CREB (Ser133) antibody (Perkin Elmer, Catalog# TRF0200) in LANCE buffer supplemented with 20 mM EDTA. The stoppedreaction mixtures were incubated at room temperature for 90 minutes. TheTR-FRET 665/620 ratios were determined using a Clariostar monochromatorplate reader instrument (BMG LabTech) in accordance with themanufacturer's instructions. The ICso value for each compound wasdetermined from each dose-response curve using GraphPad Prism 7 softwarewith a sigmodial dose response. The results for exemplary compounds ofFormula (I) and Formula (II) are shown in Table 1. Key: A is IC₅₀≤100nM; B is 100 nM<IC₅₀<1000 nM; C is 1,000<IC₅₀≤10,000 nM; D isIC₅₀>10,000 nM

TABLE 1 Inhibition of ROCK Activity by Exemplary Compounds of Formula(I) Example Number ROCK1 ROCK2 PKACA AKT1 PKG 1 C C D D D 2 D D D D D 3C C D D D 4 B B D D D 5 C C D D D 6 C B D D D 7 C C D D D 8 D D D D D 9D D D D D 10 D D D D D 11 C C D D D 12 D D D D D 13 D D D D D 14 D D D DD 15 B A D D D 16 C C D D D 17 B B D D D 18 B B D D D 19 D D D D D 20 DD D D D 21 D D D D D 22 A A C D D 23 B B D D D 24 C C D D D 25 C C D D D26 B B D D D 27 A A C C D 28 B B D D D 29 A A C C D 30 A A C C D 31 A AC D D 32 A A D D C 33 A A D D D 34 C C D D D 35 A A D D C 36 A A D D C37 B B D D D 38 B B D D D 39 B B D D D 40 A A D D D 41 B B D D D 42 B AD D D 43 C B D D D 44 C C D D D 45 A A B A C 46 N/A N/A N/A N/A N/A 47N/A N/A N/A N/A N/A 48 N/A N/A N/A N/A N/A 49 N/A N/A N/A N/A N/A 50 N/AN/A N/A N/A N/A

While the invention has been described in connection with specificembodiments thereof, it will be understood that it is capable of furthermodifications and this application is intended to cover any variations,uses, or adaptations of the invention following, in general, theprinciples of the invention and including such departures from thepresent disclosure as come within known or customary practice within theart to which the invention pertains and as may be applied to theessential features hereinbefore set forth, and as follows in the scopeof the appended claims.

We claim:
 1. A compound is selected from the group consisting of:

and pharmaceutically acceptable salts thereof.
 2. A pharmaceuticalcomposition, comprising an effective amount of a compound according toclaim 1, and a pharmaceutically acceptable excipient.
 3. A method forinhibiting ROCK activity in a cell, comprising contacting the cell inwhich inhibition of ROCK activity is desired with a therapeuticallyeffective amount of a compound of claim
 1. 4. The method of claim 3,wherein the contacting occurs in vivo.
 5. The method of claim 3, whereinthe therapeutically effective amount of the compound is between about0.01 to 300 mg/kg per day.
 6. The method of claim 5, wherein thetherapeutically effective amount of the compound is between about 0.1 to100 mg/kg per day.
 7. A method for treating a patient having cerebralcavernous malformation syndrome (CCM), comprising administering to thepatient a therapeutically effective amount of a compound of claim 1,alone or combined with a pharmaceutically acceptable carrier, exicipientor diluents.
 8. The method of claim 7, wherein the therapeuticallyeffective amount of the compound is between about 0.01 to 300 mg/kg perday.
 9. The method of claim 8, wherein the therapeutically effectiveamount of the compound is between about 0.1 to 100 mg/kg per day.
 10. Amethod of treating a cardiovascular disease associated with increasedvasotension comprising administering a therapeutically effective amountof a compound of claim 1, alone or combined with a pharmaceuticallyacceptable carrier, exicipient or diluents, to a patient in needthereof.
 11. The method of claim 10, wherein the cardiovascular diseaseis hypertension, atherosclerosis, ischemic stroke, coronary vasospasm,cerebral vasospasm, angina or erectile dysfunction.
 12. A method oftreating diseases involving elevated non-vascular smooth musclecontractility comprising administering a therapeutically effectiveamount of a compound of claim 1, alone or combined with apharmaceutically acceptable carrier, exicipient or diluents, to apatient in need thereof.
 13. The method of claim 12, wherein the diseaseinvolving elevated non-vascular smooth muscle contractility is asthma orglaucoma.